Novel compounds

ABSTRACT

Novel substituted 2, 4, 8-trisubstituted-8H-pyrido[2,3-d]pyrimidin-7-one compounds and compositions for use in therapy as CSBP/p38 kinase inhibitors.

FIELD OF THE INVENTION

This invention relates to a novel group of2,4,8-trisubstituted-8H-pyrido[2,3-d]pyrimidin-7-one compounds,processes for the preparation thereof, the use thereof in treatingCSBP/p38 kinase mediated diseases and pharmaceutical compositions foruse in such therapy.

BACKGROUND OF THE INVENTION

Intracellular signal transduction is the means by which cells respond toextracellular stimuli. Regardless of the nature of the cell surfacereceptor (e.g. protein tyrosine kinase or seven-transmembrane G-proteincoupled), protein kinases and phosphatases along with phopholipases arethe essential machinery by which the signal is further transmittedwithin the cell [Marshall, J. C. Cell, 80, 179-278 (1995)]. Proteinkinases can be categorized into five classes with the two major classesbeing, tyrosine kinases and serine/threonine kinases depending uponwhether the enzyme phosphorylates its substrate(s) on specifictyrosine(s) or serine/threonine(s) residues [Hunter, T., Methods inEnzymology (Protein Kinase Classification) p. 3, Hunter, T.; Sefton, B.M.; eds. vol. 200, Academic Press; San Diego, 1991].

For most biological responses, multiple intracellular kinases areinvolved and an individual kinase can be involved in more than onesignaling event. These kinases are often cytosolic and can translocateto the nucleus or the ribosomes where they can affect transcriptionaland translational events, respectively. The involvement of kinases intranscriptional control is presently much better understood than theireffect on translation as illustrated by the studies on growth factorinduced signal transduction involving MAP/ERK kinase [Marshall, C. J.Cell, 80, 179 (1995); Herskowitz, I. Cell , 80, 187 (1995); Hunter, T.Cell, 80, 225 (1995); Seger, R., and Krebs, E. G. FASEB J., 726-735(1995)].

While many signaling pathways are part of cell homeostasis, numerouscytokines (e.g., IL-1 and TNF) and certain other mediators ofinflammation (e.g., COX-2, and iNOS) are produced only as a response tostress signals such as bacterial lipopolysaccharide (LPS). The firstindications suggesting that the signal transduction pathway leading toLPS-induced cytokine biosynthesis involved protein kinases came fromstudies of Weinstein [Weinstein, et al., J. Immunol. 151, 3829(1993)]but the specific protein kinases involved were not identified. Workingfrom a similar perspective, Han [Han, et al., Science 265, 808(1994)]identified murine p38 as a kinase which is tyrosine phosphorylated inresponse to LPS. Definitive proof of the involvement of the p38 kinasein LPS-stimulated signal transduction pathway leading to the initiationof proinflammatory cytokine biosynthesis was provided by the independentdiscovery of p38 kinase by Lee [Lee; et al., Nature, 372, 739(1994)] asthe molecular target for a novel class of anti-inflammatory agents. Thediscovery of p38 (termed by Lee as CSBP 1 and 2) provided a mechanism ofaction of a class of anti-inflammatory compounds for which SK&F 86002was the prototypic example. These compounds inhibited IL-1 and TNFsynthesis in human monocytes at concentrations in the low uM range [Lee,et al., Int. J. Immunopharmac. 10(7), 835(1988)] and exhibited activityin animal models which are refractory to cyclooxygenase inhibitors [Lee;et al., Annals N.Y. Acad. Sci., 696, 149(1993)].

It is now firmly established that CSBP/p38 is a one of several kinasesinvolved in a stress-response signal transduction pathway which isparallel to and largely independent of the analogous mitogen-activatedprotein kinase (MAP) kinase cascade. Stress signals, including LPS,pro-inflammatory cytokines, oxidants, UV light and osmotic stress,activate kinases upstream from CSBP/p38 which in turn phosphorylateCSBP/p38 at threonine 180 and tyrosine 182 resulting in CSBP/p38activation. MAPKAP kinase-2 and MAPKAP kinase-3 have been identified asdownstream substrates of CSBP/p38 which in turn phosphorylate heat shockprotein Hsp 27 (FIG. 1). Additional downstream substrates known to bephosphorylated by p38 include kinases (Mnk1/2, MSK1/2 and PRAK) andtranscription factors (CHOP, MEF2, ATF2 and CREB). While many of thesignaling pathways required for cytokine biosynthesis remain unknown itappears clear that many of the substrates for p38 listed above areinvolved. [Cohen, P. Trends Cell Biol., 353-361(1997) and Lee, J. C. etal, Pharmacol. Ther. vol. 82, nos. 2-3, pp. 389-397, 1999].

What is known, however, is that in addition to inhibiting IL-1 and TNF,CSBP/p38 kinase inhibitors (SK&F 86002 and SB 203580) also decrease thesynthesis of a wide variety of pro-inflammatory proteins including,IL-6, IL-8, GM-CSF and COX-2. Inhibitors of CSBP/p38 kinase have alsobeen shown to suppress the TNF-induced expression of VCAM-1 onendothelial cells, the TNF-induced phosphorylation and activation ofcytosolic PLA2 and the IL-1-stimulated synthesis of collagenase andstromelysin. These and additional data demonstrate that CSBP/p38 isinvolved not only cytokine synthesis, but also in cytokine signaling[CSBP/P38 kinase reviewed in Cohen, P. Trends Cell Biol.,353-361(1997)].

Interleukin-1 (IL-1) and Tumor Necrosis Factor (TNF) are biologicalsubstances produced by a variety of cells, such as monocytes ormacrophages. IL-1 has been demonstrated to mediate a variety ofbiological activities thought to be important in immunoregulation andother physiological conditions such as inflammation [See, e.g.,Dinarello et al., Rev. Infect. Disease, 6, 51 (1984)]. The myriad ofknown biological activities of IL-1 include the activation of T helpercells, induction of fever, stimulation of prostaglandin or collagenaseproduction, neutrophil chemotaxis, induction of acute phase proteins andthe suppression of plasma iron levels.

There are many disease states in which excessive or unregulated IL-1production is implicated in exacerbating and/or causing the disease.These include rheumatoid arthritis, osteoarthritis, endotoxemia and/ortoxic shock syndrome, other acute or chronic inflammatory disease statessuch as the inflammatory reaction induced by endotoxin or inflammatorybowel disease; tuberculosis, atherosclerosis, muscle degeneration,cachexia, psoriatic arthritis, Reiter's syndrome, rheumatoid arthritis,gout, traumatic arthritis, rubella arthritis, and acute synovitis.Evidence also links IL-1 activity to diabetes and pancreatic B cells[review of the biological activities which have been attributed to IL-1Dinarello, J. Clinical Immunology, 5 (5), 287-297 (1985)].

Excessive or unregulated TNF production has been implicated in mediatingor exacerbating a number of diseases including rheumatoid arthritis,rheumatoid spondylitis, osteoarthritis, gouty arthritis and otherarthritic conditions; sepsis, septic shock, endotoxic shock, gramnegative sepsis, toxic shock syndrome, adult respiratory distresssyndrome, cerebral malaria, chronic pulmonary inflammatory disease,silicosis, pulmonary sarcoisosis, bone resorption diseases, reperfusioninjury, graft vs. host reaction, allograft rejections, fever andmyalgias due to infection, such as influenza, cachexia secondary toinfection or malignancy, cachexia, secondary to acquired immunedeficiency syndrome (AIDS), AIDS, ARC (AIDS related complex), keloidformation, scar tissue formation, Crohn's disease, ulcerative colitis,or pyresis.

Interleukin-8 (IL-8) is a chemotactic factor produced by several celltypes including mononuclear cells, fibroblasts, endothelial cells, andkeratinocytes. Its production from endothelial cells is induced by IL-1,TNF, or lipopolysachharide (LPS). IL-8 stimulates a number of functionsin vitro. It has been shown to have chemoattractant properties forneutrophils, T-lymphocytes, and basophils. In addition it induceshistamine release from basophils from both normal and atopic individualsas well as lysozomal enzyme release and respiratory burst fromneutrophils. IL-8 has also been shown to increase the surface expressionof Mac-1 (CD11b/CD18) on neutrophils without de novo protein synthesis,this may contribute to increased adhesion of the neutrophils to vascularendothelial cells. Many diseases are characterized by massive neutrophilinfiltration. Conditions associated with an increased in IL-8 production(which is responsible for chemotaxis of neutrophil into the inflammatorysite) would benefit by compounds which are suppressive of IL-8production.

IL-1 and TNF affect a wide variety of cells and tissues and thesecytokines as well as other leukocyte derived cytokines are important andcritical inflammatory mediators of a wide variety of disease states andconditions. The inhibition of these cytokines is of benefit incontrolling, reducing and alleviating many of these disease states.

In addition to the involvement of CSBP/p38 signaling in the productionof IL-1, TNF, IL-8, IL-6, GM-CSF, COX-2, collagenase and stromelysin,signal transduction via CSBP/p38 is required for the action of severalof these same pro-inflammatory proteins plus many others (VEGF, PDGF,NGF) [Ono, K. and Han, J., Cellular Signalling, 12 1-13 (2000)]. Theinvolvement of CSBP/p38 in multiple stress-induced signal transductionpathways provides additional rationale for the potential utility ofCSBP/p38 in the treatment of diseases resulting from the excessive anddestructive activation of the immune system. This expectation issupported by the potent and diverse activities described for CSBP/p38kinase inhibitors [Badger, et al., J. Pharm. Exp. Thera. 279 (3):1453-1461.(1996); Griswold, et al, Pharmacol. Comm. 7, 323-229 (1996);Jackson, et al., J. Pharmacol. Exp. Ther. 284,687-692 (1998);Underwood,et al., J. Pharmacol. Exp. Ther. 293, 281-288 (2000); Badger, et al.,Arthritis Rheum. 43, 175-183 (2000)].

There remains a need for treatment, in this field, for compounds whichare cytokine suppressive anti-inflammatory drugs, i.e. compounds whichare capable of inhibiting the CSBP/p38/RK kinase.

Other pyrido[2,3-d]pyrimidine containing pharmacophores having varyingpharmaceutical, insecticidal, and herbicidal activity may be found inthe art, such as in WO 98/33798; WO 98/23613; WO 95/19774, now U.S. Pat.No. 6,265,410; WO 00/23444; WO 01/19828 (published after the filing dateof this application); U.S. Pat. No. 5,532,370; U.S. Pat. No. 5,597,776;JP 2000-38350; WO 00/43374; WO 98/08846; and WO 01/55147 (also publishedafter the filing date of this application).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 demonstrates the p38 kinase cascade.

SUMMARY OF THE INVENTION

This invention relates to the novel compounds of Formula (I) and (Ia),and Formula (II) and (IIa), and pharmaceutical compositions comprising acompound of Formula (I) and (Ia), and Formula (II) and (IIa), and apharmaceutically acceptable diluent or carrier.

This invention relates to a method of treating a CSBP/RK/p38 kinasemediated disease in a mammal in need thereof, which comprisesadministering to said mammal an effective amount of a compound ofFormula (I) and (Ia), and Formula (II) and (IIa).

This invention also relates to a method of inhibiting cytokines and thetreatment of a cytokine mediated disease, in a mammal in need thereof,which comprises administering to said mammal an effective amount of acompound of Formula (I) and (Ia), and Formula (II) and (IIa).

This invention more specifically relates to a method of inhibiting theproduction of IL-1 in a mammal in need thereof which comprisesadministering to said mammal an effective amount of a compound ofFormula (I) and (Ia), and Formula (II) and (IIa).

This invention more specifically relates to a method of inhibiting theproduction of IL-6 in a mammal in need thereof which comprisesadministering to said mammal an effective amount of a compound ofFormula (I) and (Ia), and Formula (II) and (IIa).

This invention more specifically relates to a method of inhibiting theproduction of IL-8 in a mammal in need thereof which comprisesadministering to said mammal an effective amount of a compound ofFormula (I) and (Ia), and Formula (II) and (IIa).

This invention more specifically relates to a method of inhibiting theproduction of TNF in a mammal in need thereof which comprisesadministering to said mammal an effective amount of a compound ofFormula (I) and (Ia), and Formula (II) and (IIa).

Accordingly, the present invention provides a compound of Formula (I)and (Ia):

wherein

-   R₁ is an optionally substituted aryl or an optionally substituted    heteroaryl ring;-   R₂ is hydrogen, C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, C₃₋₇ cycloalkylalkyl,    aryl, arylC₁₋₁₀ alkyl, heteroaryl, heteroarylC₁₋₁₀ alkyl,    heterocyclic, or a heterocyclylC₁₋₁₀ alkyl moiety, which moieties    are all optionally substituted, or R₂ is the moiety    X₁(CR₁₀R₂₀)_(q)C(A₁)(A₂)(A₃), or C(A₁)(A₂)(A₃);-   A₁ is an optionally substituted C₁₋₁₀ alkyl;-   A₂ is an optionally substituted C₁₋₁₀ alkyl;-   A₃ is hydrogen or is an optionally substituted C₁₋₁₀ alkyl;-   R₃ is an C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, C₃₋₇ cycloalkylC₁₋₄alkyl,    aryl, arylC₁₋₁₀ alkyl, heteroaryl, heteroarylC₁₋₁₀ alkyl,    heterocyclic, or a heterocyclylC₁₋₁₀ alkyl moiety, which moieties    are optionally substituted;-   R₄ and R₁₄ are each independently selected from hydrogen, optionally    substituted C₁₋₄ alkyl, optionally substituted C₃₋₇ cycloalkyl, C₃₋₇    cycloalkylC₁₋₄alkyl, optionally substituted aryl, or optionally    substituted aryl-C₁₋₄ alkyl, or R₄ and R₁₄ together with the    nitrogen which they are attached form an optionally substituted    heterocyclic ring of 4 to 7 members, which ring optionally contains    an additional heteroatom selected from oxygen, sulfur or NR₉;-   R₆ is hydrogen, C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, heterocyclyl,    heterocyclyl C₁₋₁₀alkyl, aryl, arylC₁₋₁₀ alkyl, heteroaryl or    heteroarylC₁₋₁₀ alkyl, wherein each of these moieties may be    optionally substituted;-   R₉ is hydrogen, C(Z)R₆ or optionally substituted C₁₋₁₀ alkyl,    optionally substituted aryl or optionally substituted aryl-C₁₋₄    alkyl;-   R₁₀ and R₂₀ are independently selected from hydrogen or C₁₋₄alkyl;-   X is R₂, OR₂, S(O)_(m)R₂, (CH₂)_(n)N(R₁₀)S(O)_(m)R₂,    (CH₂)_(n)N(R₁₀)C(O)R₂, (CH₂)_(n)NR₄R₁₄, or (CH₂)_(n)N(R₂)₂;-   X₁ is N(R₁₀), O, S(O)_(m), or CR₁₀R₂₀;-   n is 0 or an integer having a value of 1 to 10;-   m is 0 or an integer having a value of 1 or 2;-   q is 0 or an integer having a value of 1 to 10;-   Z is oxygen or sulfur;    or a pharmaceutically acceptable salt thereof.

DETAILED DESCRIPTION OF THE INVENTION

Another aspect of the present invention provides for the compound ofFormula (II) and (IIa):

wherein

-   R₁ is the moiety YRa;-   R₂ is hydrogen, C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, C₃₋₇ cycloalkylalkyl,    aryl, arylC₁₋₁₀ alkyl, heteroaryl, heteroarylC₁₋₁₀ alkyl,    heterocyclic, or a heterocyclylC₁₋₁₀ alkyl moiety, which moieties    are all optionally substituted, or R₂ is the moiety X₁(CR₁₀R₂₀)_(q)    C(A₁)(A₂)(A₃), or C(A₁)(A₂)(A₃);-   A₁ is an optionally substituted C₁₋₁₀ alkyl;-   A₂ is an optionally substituted C₁₋₁₀ alkyl;-   A₃ is hydrogen or is an optionally substituted C₁₋₁₀ alkyl;-   R₃ is an C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, C₃₋₇ cycloalkylC₁₋₄alkyl,    aryl, arylC₁₋₁₀ alkyl, heteroaryl, heteroarylC₁₋₁₀ alkyl,    heterocyclic, or a heterocyclylC₁₋₁₀ alkyl moiety, which moieties    are optionally substituted;-   R₄ and R₁₄ are each independently selected from hydrogen, optionally    substituted C₁₋₄ alkyl, optionally substituted C₃₋₇ cycloalkyl, C₃₋₇    cycloalkylC₁₋₄alkyl, optionally substituted aryl, or optionally    substituted aryl-C₁₋₄ alkyl, or R₄ and R₁₄ together with the    nitrogen which they are attached form an optionally substituted    heterocyclic ring of 4 to 7 members, which ring optionally contains    an additional heteroatom selected from oxygen, sulfur or NR₉;-   R₆ is hydrogen, C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, heterocyclyl,    heterocyclyl C₁₋₁₀alkyl, aryl, arylC₁₋₁₀ alkyl, heteroaryl or    heteroarylC₁₋₁₀ alkyl, wherein each of these moieties may be    optionally substituted;-   R₉ is hydrogen, C(Z)R₆ or optionally substituted C₁₋₁₀ alkyl,    optionally substituted aryl or optionally substituted aryl-C₁₋₄    alkyl;-   R₁₀ and R₂₀ are independently selected from hydrogen or C₁₋₄alkyl;-   Y is C(R_(b))(R_(d)), C(O), N(R_(d)), N(R_(d))C(R_(c))(R_(d)),    oxygen, OC(R_(c))(R_(d)), S(O)m, or S(O)_(m)C(R_(c))(R_(d));-   R_(a) is an aryl or heteroaryl ring, which ring is optionally    substituted;-   R_(b) is hydrogen, C₁₋₂ alkyl, NR_(c), hydroxy, thio, C₁₋₂ alkoxy,    S(O)_(m)C₁₋₂ alkyl;-   R_(c) is hydrogen or C₁₋₂ alkyl;-   R_(d) is hydrogen or C₁₋₂ alkyl;-   X is R₂, OR₂, S(O)_(m)R₂, (CH₂)_(n)N(R₁₀)S(O)_(m)R₂,    (CH₂)_(n)N(R₁₀)C(O)R₂, (CH₂)_(n)NR₄R₁₄, or (CH₂)_(n)N(R₂)₂;-   X₁ is N(R₁₀), O, S(O)_(m), or CR₁₀R₂₀;-   n is 0 or an integer having a value of 1 to 10;-   m is 0 or an integer having a value of 1 or 2;-   q is 0 or an integer having a value of 1 to 10;-   Z is oxygen or sulfur;    or a pharmaceutically acceptable salt thereof.

The present invention is directed to novel compounds of Formula (I) and(Ia), and those of Formula (II) and (IIa), or a pharmaceuticallyacceptable salt thereof. As will be readily recognized, the differencebetween compounds of Formula (I) and (Ia) and that of Formula (II) and(IIa) lies in the unsaturation of the pyrido-7-one ring. The respectiveR₁, R₂, X and R₃ terms are the same for both groups within the Formulaitself, for instance I and Ia. For purposes herein, everythingapplicable to Formula (I) is also applicable to Formula (Ia) unlessotherwise indicated, and everything applicable to Formula (II) is alsoapplicable to Formula (IIa) unless otherwise indicated.

Suitably, for compounds of Formula (I), and (Ia), R₁ is an aryl, orheteroaryl ring, which ring is optionally substituted. The R₁ aryl orheteroaryl rings may be substituted one or more times, preferably 1 to 4times, independently, by substituents selected from halogen, C₁₋₄ alkyl,halo-substituted-C₁₋₄ alkyl, cyano, nitro, (CR₁₀R₂₀)_(v)NR₄R₁₄,(CR₁₀R₂₀)_(v)C(Z)NR₄R₁₄, (CR₁₀R₂₀)_(v)C(Z)OR₈, (CR₁₀R₂₀)_(v)COR_(a′),(CR₁₀R₂₀)_(v)C(O)H, SR₅, S(O)R₅, S(O)₂R₅, (CR₁₀R₂₀)_(v)OR₈, ZC(Z)R₁₁,NR₁₀C(Z)R₁₁, or NR₁₀S(O)₂R₇.

Preferably, R₁ is an aryl moiety, more preferably a phenyl ring,optionally substituted one or more times by halogen, C₁₋₄ alkyl, orhalo-substituted-C₁₋₄ alkyl. More preferably, the phenyl ring issubstituted in the 2, 4, or 6-position, or di-substituted in the 2,4-position, such as 2-fluoro, 4-fluoro, 2,4-difluoro, or2-methyl-4-fluoro; or tri-substituted in the 2,4,6-position such as2,4,6-trifluoro. Another preferred embodiment is substitution of thephenyl ring in the 3-position, such as with a halogen derivative,producing a 3-position, 2,3-disubstitution, or a 3,4-disubstitution.

Preferably, when R₁ is a heteroaryl moiety, the ring is not attached tothe pharmacophore via one of the heteroatoms, such as nitrogen to form acharged ring. For instance, a pyridinyl ring would be attached through acarbon atom to yield a 2-, 3- or 4-pyridyl moiety, which is optionallysubstituted.

Suitably, v is 0 or an integer having a value of 1 or 2.

Suitably, Z is oxygen or sulfur.

Suitably, R_(a′), is C₁₋₄ alkyl, halo-substituted C₁₋₄ alkyl, C₂₋₄alkenyl, C₂₋₄ alkynyl, C₃₋₇ cycloalkyl, C₅₋₇ cycloalkenyl, aryl,arylC₁₋₄ alkyl, heteroaryl, heteroarylC₁₋₄ alkyl, heterocyclyl,heterocyclylC₁₋₄ alkyl, (CR₁₀R₂₀)_(v)OR₇, (CR₁₀R₂₀)_(v)S(O)_(m)R₇,(CR₁₀R₂₀)_(v)NR₁₀S(O)₂R₇, or (CR₁₀R₂₀)_(v)NR₄R₁₄; and wherein the aryl,arylalkyl, heteroaryl, heteroaryl alkyl may be optionally substituted.

Suitably, for compounds of Formula (II), and (IIa), R₁ is Y-R_(a).

Suitably, Y is C(R_(b))(R_(d)), C(O), N(R_(d)), N(R_(d))C(R_(c))(R_(d)),oxygen, OC(R_(c))(R_(d)), S(O)m, or S(O)_(m)C(R_(c))(R_(d)).

Suitably, R_(b) is hydrogen, C₁₋₂ alkyl, NR_(c), hydroxy, thio, C₁₋₂alkoxy, S(O)_(m)C₁₋₁₂ alkyl.

Suitably, R_(c) is hydrogen or C₁₋₂ alkyl.

Suitably, R_(d) is hydrogen or C₁₋₂ alkyl.

Suitably, m is 0 or an integer having a value of 1 or 2.

Suitably R_(a) is an optionally substituted aryl ring or an optionallysubstituted heteroaryl ring. The optional substitutents for these ringsare the same as for the Formula (I) and (Ia) R₁ aryl and heteroarylrings as noted above.

As will be appreciated the difference between compounds of Formula (I)and (II) lies in the R₁ substitution. The remaining substituent groupsare the same and for purposes herein applicable to all four formulasunless otherwise indicated.

Suitably, R₄ and R₁₄ are each independently selected from hydrogen,optionally substituted C₁₋₄ alkyl, optionally substitutedC₃₋₇cycloalkyl, optionally substituted C₃₋₇cycloalkylC₁₋₄ alkyl,optionally substituted aryl or optionally substituted aryl-C₁₋₄ alkyl,or R₄ and R₁₄ together with the nitrogen to which they are attached mayform an optionally substituted heterocyclic ring of 4 to 7 members whichring optionally contains an additional heteroatom selected from oxygen,sulfur or NR₉.

The C₁₋₄ alkyl, C₃₋₇cycloalkyl, C₃₋₇cycloalkylC₁₋₄ alkyl, aryl andaryl-C₁₋₄ alkyl moieties may be optionally substituted, one or moretimes, preferably 1 to 4 times independently by halogen, such asfluorine, chlorine, bromine or iodine; hydroxy; hydroxy substitutedC₁₋₁₀alkyl; C₁₋₁₀ alkoxy, such as methoxy or ethoxy; halosubstitutedC₁₋₁₀ alkoxy; S(O)m alkyl, such as methyl thio, methylsulfinyl or methylsulfonyl; aldehydes (—C(O)), or a ketone, such as —C(O)R₆, such asC(O)C₁₋₁₀alkyl or C(O)aryl; amides, such as C(O)NR_(4′)R_(14′), orNR_(4′)C(O)C₁₋₁₀alkyl, or NR_(4′)C(O)aryl; NR_(4′)R_(14′), whereinR_(4′) and R_(14′) are each independently hydrogen or C₁₋₄ alkyl, orwherein the R_(4′)R_(14′) can cyclize together with the nitrogen towhich they are attached to form a 5 to 7 membered ring which optionallycontains an additional heteroatom selected from O/N/S; cyano, nitro,C₁₋₁₀ alkyl, C₃₋₇cycloalkyl, or C₃₋₇cycloalkyl C₁₋₁₀ alkyl group, suchas methyl, ethyl, propyl, isopropyl, t-butyl, etc. or cyclopropylmethyl; halosubstituted C₁₋₁₀ alkyl, such CF₂CF₂H, CH₂CF₃, or CF₃; anoptionally substituted aryl, such as phenyl, or an optionallysubstituted arylalkyl, such as benzyl or phenethyl, wherein these arylcontaining moieties may also be substituted one to two times by halogen;hydroxy; hydroxy substituted alkyl; C₁₋₁₀ alkoxy; S(O)_(m)alkyl; amino,mono & di-substituted C₁₋₄ alkyl amino, such as in the NR_(4′)R_(14′)group; C₁₋₄ alkyl, or CF₃.

When R₄ and R₁₄ together with the nitrogen cyclize to form a ring,suitably, such rings include, but are not limited to pyrrolidine,piperidine, piperazine, morpholine, and thiomorpholine (includingoxidizing the sulfur). The ring may be optional substituted, one or moretimes, preferably 1 to 4 times, independently by halogen, such asfluorine, chlorine, bromine or iodine; hydroxy; hydroxy substitutedC₁₋₁₀alkyl; C₁₋₁₀ alkoxy, such as methoxy or ethoxy; halosubstitutedC₁₋₁₀ alkoxy; S(O)m alkyl, such as methyl thio, methylsulfinyl or methylsulfonyl; a ketone on the cyclized ring (—C(O)), or a ketone or aldehydeoff the ring (—C(O)R₆), such as C(O)C₁₋₁₀ alkyl or C(O) aryl;NR_(4′)R_(14′), wherein R_(4′) and R_(14′) are each independentlyhydrogen or C₁₋₄ alkyl; C₁₋₁₀ alkyl, C₃₋₇cycloalkyl, or C₃₋₇cycloalkylC₁₋₁₀ alkyl group, such as methyl, ethyl, propyl, isopropyl, t-butyl,etc. or cyclopropyl methyl; halosubstituted C₁₋₁₀ alkyl, such CF₂CF₂H,CH₂CF₃, or CF₃; an optionally substituted aryl, such as phenyl, or anoptionally substituted arylalkyl, such as benzyl or phenethyl, whereinthese aryl containing moieties may also be substituted one to two timesby halogen; hydroxy; hydroxy substituted alkyl; C₁₋₁₀ alkoxy;S(O)_(m)alkyl; amino, mono & di-substituted C₁₋₄ alkyl amino, such as inthe NR_(4′)R_(14′) group; C₁₋₄ alkyl, or CF₃.

Suitably, R₅ is hydrogen, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl orNR₄R₁₄, excluding the moieties SR₅ being SNR₄R₁₄, S(O)₂R₅ being SO₂H andS(O)R₅ being SOH.

Suitably, R₆ is hydrogen, C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, heterocyclyl,heterocyclyl C₁₋₁₀alkyl, aryl, arylC₁₋₁₀ alkyl, heteroaryl orheteroarylC₁₋₁₀ alkyl, wherein these moieties may be optionallysubstituted.

Suitably, R₇ is C₁₋₆alkyl, aryl, arylC₁₋₆alkyl, heterocyclic,heterocyclylC₁₋₆ alkyl, heteroaryl, or heteroarylC₁₋₆alkyl; and whereineach of these moieties may be optionally substituted.

Suitably, R₈ is hydrogen, C₁₋₄ alkyl, halo-substituted C₁₋₄ alkyl, C₂₋₄alkenyl, C₂₋₄ alkynyl, C₃₋₇ cycloalkyl, C₅₋₇ cycloalkenyl, aryl,arylC₁₋₄ alkyl, heteroaryl, heteroarylC₁₋₄ alkyl, heterocyclyl,heterocyclylC₁₋₄ alkyl, (CR₁₀R₂₀)_(t)OR₇, (CR₁₀R₂₀)_(t)S(O)_(m)R₇,(CR₁₀R₂₀)_(t) NR₁₀S(O)₂R₇, or (CR₁₀R₂₀)_(t)NR₄R₁₄; and wherein thecycloalkyl, cycloalkenyl, aryl, arylalkyl, heteroaryl, heteroaryl alkyl,heterocyclic and heterocyclic alkyl moieties may be optionallysubstituted.

Suitably, t is an integer having a value of 1 to 3.

Suitably, R₉ is hydrogen, C(Z)R₆, optionally substituted C₁₋₁₀ alkyl,optionally substituted aryl or optionally substituted aryl-C₁₋₄ alkyl.

Suitably, R₁₀ and R₂₀ are independently selected from hydrogen or a C₁₋₄alkyl.

Suitably, R₁₁ is C₁₋₄ alkyl, halo-substituted C₁₋₄ alkyl, C₂₋₄ alkenyl,C₂₋₄ alkynyl, C₃₋₇ cycloalkyl, C₅₋₇ cycloalkenyl, aryl, arylC₁₋₁₄ alkyl,heteroaryl, heteroarylC₁₋₄ alkyl, heterocyclyl, heterocyclylC₁₋₄ alkyl,(CR₁₀R₂₀)_(t)OR₇, (CR₁₀R₂₀)_(t)S(O)_(m)R₇, (CR₁₀R₂₀)_(t)NR₁₀S(O)₂R₇, or(CR₁₀R₂₀)_(v)NR₄R₁₄; and wherein the aryl, arylalkyl, heteroaryl,heteroaryl alkyl, heterocyclyl, and heterocyclylalkyl moieties may beoptionally substituted.

Suitably m is 0 or an integer having a value of 1 or 2.

Suitably, R₃ is an optionally substituted C₁₋₁₀alkyl, C₃₋₇ cycloalkyl,C₃₋₇ cycloalkyl C₁₋₁₀alkyl, aryl, arylC₁₋₁₀ alkyl, heteroarylC₁₋₁₀alkyl,or heterocyclylC₁₋₁₀ alkyl moiety, which moieties are optionallysubstituted one or more times, preferably 1 to 4 times, independently byC₁₋₁₀ alkyl, halo-substituted C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl,C₃₋₇cycloalkyl, C₃₋₇cycloalkylC₁₋₁₀ alkyl, C₅₋₇cycloalkenyl,C₅₋₇cycloalkenylC₁₋₁₀ alkyl, halogen, cyano, nitro, (CR₁₀R₂₀)_(n)OR₆,(CR₁₀R₂₀)_(n)SH, (CR₁₀R₂₀)_(n)S(O)_(m)R₇, (CR₁₀R₂₀)_(n)NR₁₀S(O)₂R₇,(CR₁₀R₂₀)_(n)NR₄R₁₄, (CR₁₀R₂₀)_(n)CN, (CR₁₀R₂₀)_(n)S(O)₂NR₄R₁₄,(CR₁₀R₂₀)_(n)C(Z)R₆, (CR₁₀R₂₀)_(n)OC(Z)R₆, (CR₁₀R₂₀)_(n)C(Z)OR₆,(CR₁₀R₂₀)_(n)C(Z)NR₄R₁₄, (CR₁₀R₂₀)_(n)NR₁₀C(Z)R₆,(CR₁₀R₂₀)_(n)NR₁₀C(═NR₁₀)NR₄R₁₄, (CR₁₀R₂₀)_(n)OC(Z)NR₄R₁₄,(CR₁₀R₂₀)_(n)NR₁₀C(Z)NR₄R₁₄, or (CR₁₀R₂₀)_(n)NR₁₀C(Z)OR₇.

Preferably the optional substituents are independently selected fromhalogen, alkyl, hydroxy, alkoxy, cyano, nitro, amino, or halosubstitutedalkyl. More preferably, halogen, or alkyl.

Preferably, R₃ is an optionally substituted C₁₋₁₀ alkyl, C₃₋₇cycloalkyl,C₃₋₇cycloalkylalkyl, or aryl. More preferably, R₃ is an optionallysubstituted C₁ alkyl, or aryl.

Preferably, when R₃ is an aryl moiety, it is a phenyl ring, optionallysubstituted one or more times by halogen, C₁₋₄ alkyl, orhalo-substituted-C₁₋₄ alkyl. More preferably, the phenyl ring issubstituted in the 2, 4, or 6-position, or di-substituted in the 2,4-position, such as 2-fluoro, 4-fluoro, 2,4-difluoro, or2-methyl-4-fluoro; or tri-substituted in the 2,4,6-position, such as2,4,6-trifluoro.

Suitably, n is 0, or an integer having a value of 1 to 10.

Suitably, X is R₂, OR₂, S(O)_(m)R₂, (CH₂)_(n)N(R₁₀)S(O)mR₂,(CH₂)_(n)N(R₁₀)C(O)R₂, (CH₂)_(n)NR₄R₁₄, or (CH₂)_(n)N(R₂)₂. Preferably Xis R₂, OR₂, (CH₂)_(n)NR₄R₁₄, or (CH₂)_(n)N(R₂)₂. Preferably, when X isR₂, then R₂ is the moiety X₁(CR₁₀R₂₀)_(q)C(A₁)(A₂)(A₃), orC(A₁)(A₂)(A₃).

Suitably, R₂ is independently selected from hydrogen, optionallysubstituted C₁₋₁₀ alkyl, optionally substituted C₃₋₇ cycloalkyl,optionally substituted C₃₋₇cycloalkylalkyl, optionally substituted aryl,optionally substituted arylC₁₋₁₀alkyl, optionally substitutedheteroaryl, optionally substituted heteroarylC₁₋₁₀ alkyl, optionallysubstituted heterocyclic, optionally substituted heterocyclylC₁₋₁₀ alkylmoiety, or R₂ is the moiety X₁(CR₁₀R₂₀)_(q)C(A₁)(A₂)(A₃), orC(A₁)(A₂)(A₃).

The R₂ moieties, excluding hydrogen, may be optionally substituted oneor more times, preferably 1 to 4 times, independently by C₁₋₁₀ alkyl,halo-substituted C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₇cycloalkyl, C₃₋₇cycloalkylC₁₋₁₀alkyl, C₅₋₇cycloalkenyl, C₅₋₇cycloalkenyl C₁₋₁₀ alkyl, halogen, —C(O), cyano, nitro,(CR₁₀R₂₀)_(n)OR₆, (CR₁₀R₂₀)_(n)SH, (CR₁₀R₂₀)_(n)S(O)_(m)R₇,(CR₁₀R₂₀)_(n)NR₁₀S(O)₂R₇, (CR₁₀R₂₀)_(n)NR₄R₁₄, (CR₁₀R₂₀)_(n)CN,(CR₁₀R₂₀)_(n)S(O)₂NR₄R₁₄, (CR₁₀R₂₀)_(n)C(Z)R₆, (CR₁₀R₂₀)_(n)OC(Z)R₆,(CR₁₀R₂₀)_(n)C(Z)OR₆, (CR₁₀R₂₀)_(n)C(Z)NR₄R₁₄, (CR₁₀R₂₀)_(n)NR₁₀C(Z)R₆,(CR₁₀R₂₀)_(n)NR₁₀C(═NR₁₀)NR₄R₁₄, (CR₁₀R₂₀)_(n)C(═NOR₆)NR₄R₁₄,(CR₁₀R₂₀)_(n)OC(Z)NR₄R₁₄, (CR₁₀R₂₀)_(n)NR₁₀C(Z)NR₄R₁₄, or(CR₁₀R₂₀)_(n)NR₁₀C(Z)OR₇.

Suitably X₁ is N(R₁₀), O, S(O)_(m), or CR₁₀R₂₀. More preferably, X₁ isN(R₁₀), or 0.

Suitably, q is 0 or an integer having a value of 1 to 10.

Suitably, A₁ is an optionally substituted C₁₋₁₀ alkyl.

Suitably, A₂ is an optionally substituted C₁₋₁₀ alkyl.

Suitably, A₃ is hydrogen or is an optionally substituted C₁₋₁₀ alkyl.

The A₁, A₂, and A₃ C₁₋₁₀ alkyl moieties may optionally substituted oneor more times, independently, preferably from 1 to 4 times, withhalogen, such as chlorine, fluorine, bromine, or iodine;halo-substituted C₁₋₁₀alkyl, such as CF₃, or CHF₂CF₃; C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, C₃₋₇ cycloalkyl, C₃₋₇cycloalkylC₁₋₁₀alkyl,C₅₋₇cycloalkenyl, C₅₋₇ cycloalkenylC₁₋₁₀alkyl, (CR₁₀R₂₀)_(n)OR₆,(CR₁₀R₂₀)_(n)SH, (CR₁₀R₂₀)_(n)S(O)_(m)R₇, (CR₁₀R₂₀)_(n)NR₁₀S(O)₂R₇,(CR₁₀R₂₀)_(n)NR₄R₁₄, (CR₁₀R₂₀)_(n)CN, (CR₁₀R₂₀)_(n)S(O)₂NR₄R₁₄,(CR₁₀R₂₀)_(n)C(Z)R₆, (CR₁₀R₂₀)_(n)OC(Z)R₆, (CR₁₀R₂₀)_(n)C(Z)OR₆,(CR₁₀R₂₀)_(n)C(Z)NR₄R₁₄, (CR₁₀R₂₀)_(n)NR₁₀C(Z)R₆,(CR₁₀R₂₀)_(n)NR₁₀C(═NR₁₀)NR₄R₁₄, (CR₁₀R₂₀)_(n)OC(Z)NR₄R₁₄,(CR₁₀R₂₀)_(n)NR₁₀C(Z)NR₄R₁₄, or (CR₁₀R₂₀)_(n)NR₁₀C(Z)OR₇.

Preferably, one or more of A₁ to A₃ is substituted with(CR₁₀R₂₀)_(n)OR₆More preferably, R₆ is hydrogen.

A preferred C(A₁)(A₂)(A₃) grouping is CH(CH₂OH)₂, or C(CH₃)(CH₂OH)₂,X₁(CR₁₀R₂₀)_(q)CH(CH₂OH)₂, or X₁(CR₁₀R₂₀)_(q)C(CH₃)(CH₂OH)₂. X₁ ispreferably oxygen or nitrogen.

As used herein, “optionally substituted” unless specifically definedshall mean such groups as halogen, such as fluorine, chlorine, bromineor iodine; hydroxy; hydroxy substituted C₁₋₁₀alkyl; C₁₋₁₀ alkoxy, suchas methoxy or ethoxy; halosubstituted C₁₋₁₀ alkoxy; S(O)m alkyl, such asmethyl thio, methylsulfinyl or methyl sulfonyl; —C(O); NR_(4′)R_(14′),wherein R_(4′) and R_(14′) are each independently hydrogen or C₁₋₄alkyl, such as amino or mono or -disubstituted C₁₋₄ alkyl or wherein theR_(4′)R_(14′) can cyclize together with the nitrogen to which they areattached to form a 5 to 7 membered ring which optionally contains anadditional heteroatom selected from O/N/S; C₁₋₁₀ alkyl, C₃₋₇cycloalkyl,or C₃₋₇cycloalkyl C₁₋₁₀ alkyl group, such as methyl, ethyl, propyl,isopropyl, t-butyl, etc. or cyclopropyl methyl; halosubstituted C₁₋₁₀alkyl, such CF₂CF₂H, or CF₃; an optionally substituted aryl, such asphenyl, or an optionally substituted arylalkyl, such as benzyl orphenethyl, wherein these aryl containing moieties may also besubstituted one to two times by halogen; hydroxy; hydroxy substitutedalkyl; C₁₋₁₀ alkoxy; S(O)_(m)alkyl; amino, mono & di-substituted C₁₋₄alkyl amino, such as in the NR₄R₁₄ group; C₁₋₄ alkyl, or CF₃.

Suitable pharmaceutically acceptable salts are well known to thoseskilled in the art and include basic salts of inorganic and organicacids, such as hydrochloric acid, hydrobromic acid, sulphuric acid,phosphoric acid, methane sulphonic acid, ethane sulphonic acid, aceticacid, malic acid, tartaric acid, citric acid, lactic acid, oxalic acid,succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid,phenylacetic acid and mandelic acid.

In addition, pharmaceutically acceptable salts of compounds of Formula(I) may also be formed with a pharmaceutically acceptable cation, forinstance, if a substituent group comprises a carboxy moiety. Suitablepharmaceutically acceptable cations are well known to those skilled inthe art and include alkaline, alkaline earth, ammonium and quaternaryammonium cations.

The term “halo” or “halogens” is used herein to mean the halogens,chloro, fluoro, bromo and iodo.

The term “C₁₋₁₀alkyl” or “alkyl” or “alkyl₁₋₁₀” is used herein to meanboth straight and branched chain radicals of 1 to 10 carbon atoms,unless the chain length is otherwise limited, including, but not limitedto, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl,tert-butyl, n-pentyl and the like.

The term “cycloalkyl” is used herein to mean cyclic radicals, preferablyof 3 to 8 carbons, including but not limited to cyclopropyl,cyclopentyl, cyclohexyl, and the like.

The term “cycloalkenyl” is used herein to mean cyclic radicals,preferably of 5 to 8 carbons, which have at least one bond including butnot limited to cyclopentenyl, cyclohexenyl, and the like.

The term “alkenyl” is used herein at all occurrences to mean straight orbranched chain radical of 2-10 carbon atoms, unless the chain length islimited thereto, including, but not limited to ethenyl, 1-propenyl,2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl and the like.

The term “aryl” is used herein to mean phenyl and naphthyl.

The term “heteroaryl” (on its own or in any combination, such as“heteroaryloxy”, or “heteroaryl alkyl”) is used herein to mean a 5-10membered aromatic ring system in which one or more rings contain one ormore heteroatoms selected from the group consisting of N, O or S, suchas, but not limited, to pyrrole, pyrazole, furan, pyran, thiophene,quinoline, isoquinoline, quinazolinyl, pyridine, pyrimidine, pyridazine,pyrazine, uracil, oxadiazole, oxazole, isoxazole, oxathiadiazole,thiazole, isothiazole, thiadiazole, tetrazole, triazole, indazole,imidazole, or benzimidazole.

The term “heterocyclic” (on its own or in any combination, such as“heterocyclylalkyl”) is used herein to mean a saturated or partiallyunsaturated 4-10 membered ring system in which one or more rings containone or more heteroatoms selected from the group consisting of N, O, S,or S(O)m, and m is 0 or an integer having a value of 1 or 2; such as,but not limited to, the saturated or partially saturated versions of theheteroaryl moieties as defined above, such as tetrahydropyrrole,tetrahydropyran, tetrahydrofuran, tetrahydrothiophene (includingoxidized versions of the sulfur moiety), pyrrolidine, piperidine,piperazine, morpholine, thiomorpholine (including oxidized versions ofthe sulfur moiety), or imidazolidine.

The term “aralkyl” or “heteroarylalkyl” or “heterocyclicalkyl” is usedherein to mean C₁₋₄ alkyl as defined above attached to an aryl,heteroaryl or heterocyclic moiety as also defined herein unlessotherwise indicate.

The term “sulfinyl” is used herein to mean the oxide S(O) of thecorresponding sulfide, the term “thio” refers to the sulfide, and theterm “sulfonyl” refers to the fully oxidized S(O)₂ moiety.

The term “aroyl” is used herein to mean C(O)Ar, wherein Ar is as phenyl,naphthyl, or aryl alkyl derivative such as defined above, such groupinclude but are not limited to benzyl and phenethyl.

The term “alkanoyl” is used herein to mean C(O)C₁₋₁₀ alkyl wherein thealkyl is as defined above.

It is recognized that the compounds of the present invention may existas stereoisomers, regioisomers, or diastereiomers. These compounds maycontain one or more asymmetric carbon atoms and may exist in racemic andoptically active forms. All of these individual compounds, isomers, andmixtures thereof are included within the scope of the present invention.

Exemplified compounds of the compounds of this invention include theracemates, or optically active forms of the compounds of the workingexamples herein, and pharmaceutically acceptable salts thereof.

Methods of Manufacture

The compounds of Formula (I), (Ia), (II) and (IIa) may be obtained byapplying synthetic procedures, described herein. The synthesis providedfor is applicable to producing compounds of Formula (I), (Ia), (II) and(IIa) having a variety of different R₁, R₂, Y, X, and R₃ groups whichare reacted, employing optional substituents which are suitablyprotected, to achieve compatibility with the reactions outlined herein.Subsequent deprotection, in those cases, then affords compounds of thenature generally disclosed. While a particular formula with particularsubstituent groups is shown herein, the synthesis is applicable to allformulas and all substituent groups herein.

Once the nucleus has been established, further compounds of Formula (I),(Ia), (II) and (IIa) may be prepared by applying standard techniques forfunctional group interconversion, well known in the art. For instance:C(O)NR₄R₁₄ from CO₂CH₃ by heating with HNR₄R₁₄ in CH₃OH with or withoutcatalytic or stoichiometric metal cyanide or Aluminum trimethyl, e.g.NaCN; OC(O)R₃ from OH with e.g., ClC(O)R₆ in bases such as triethylamineand pyridine; NR₁₀—C(S)NR₄R₁₄ from NHR₁₀ with an alkylisothiocyanate, orthiocyanic acid and ClC(S)NR₄R₁₄; NR₁₀C(O)OR₆ from NHR₁₀ with an alkylor aryl chloroformate; NR₁₀C(O)NR₄H from NHR₁₀ by treatment with anisocyanate, e.g. R₄N═C═O; NR₁₀—C(O)R₆ from NHR₁₀ by treatment withCl—C(O)R₆ in pyridine; C(═NR₁₀)NR₄R₁₄ from C(NR₄R₁₄)S with H₃NR₁₀ ⁺OAc⁻by heating in alcohol; C(NR₄R₁₄)SR₆ from C(S)NR₄R₁₄ with R₆-I in aninert solvent, e.g. acetone; NR₁₀SO₂R₇ from NHR₁₀ by treatment withClSO₂R₇ by heating in bases such as pyridine; NR₁₀C(S)R₆ from NR₁₀C(O)R₆by treatment with Lawesson's reagent[2,4-bis(4-methoxyphenyl)-1,3,2,4dithiadiphosphetane-2,4-disulfide];NR₁₀SO₂CF₃ from NHR₁₀ with triflic anhydride and base wherein R₃, R₆,R₁₀, R₄ and R₁₄ are as defined in Formula (I) herein.

Precursors of the groups R₁, R₂ and R₃, can be other R₁, R₂ and R₃, etc.groups that may be interconverted by applying standard techniques forfunctional group interconversion. For example wherein a moiety is a halosubstituted C₁₋₁₀ alkyl can be converted to the corresponding C₁₋₁₀alkylN₃ derivative by reacting with a suitable azide salt, andthereafter if desired can be reduced to the corresponding C₁₋₁₀alkylNH₂compound, which in turn can be reacted with R₇S(0)₂X wherein X is halo(e.g., chloro) to yield the corresponding C₁₋₁₀alkylNHS(0)₂R₇ compound.

Alternatively wherein the moiety is a halo-substituted C₁₋₁₀-alkyl itcan be reacted with an amine R₄R₁₄NH to yield the correspondingC₁₋₁₀-alkylNR₄R₁₄ compound, or can be reacted with an alkali metal saltof R₇SH to yield the corresponding C₁₋₁₀alkylSR₇ compound.

As noted above, it may be desirable during the synthesis of thecompounds of this invention, to derivatize reactive functional groups inthe molecule undergoing reaction so as to avoid unwanted side reactions.Functional groups such as hydroxy, amino, an acid groups typically areprotected with suitable groups that can be readily removed when desired.Suitable common protecting groups for use with hydroxyl groups andnitrogen groups are well known in the art and described in manyreferences, for instance, Protecting Groups in Organic Synthesis, Greeneet al., John Wiley & Sons, New York, N.Y., (2nd edition, 1991 or theearlier 1981 version). Suitable examples of hydroxyl protecting groupsinclude ether forming groups such as benzyl, and aryl groups such astert-butoxycarbonyl (Boc), silyl ethers, such as t-butyldimethyl ort-butyldiphenyl, and alkyl ethers, such as methyl connected by an alkylchain of variable link, (CR₁₀R₂₀)_(n). Amino protecting groups mayinclude benzyl, aryl such as acetyl and trialkylsilyl groups. Carboxylicacid groups are typically protected by conversion to an ester that caneasily be hydrolyzed, for example, trichloethyl, tert-butyl, benzyl andthe like.

Pharmaceutically acid addition salts of compounds of Formula (I), (Ia),(II) and (IIa) may be obtained in known manner, for example by treatmentthereof with an appropriate amount of acid in the presence of a suitablesolvent.

An illustration of the preparation of compounds of the present inventionis shown in the scheme below. For purposes herein, the compounds inSchemes I and II are shown with an S-methyl, or S(O)₂-methyl group whichis deemed representative of the S(O)m-Rg group, as described in theformulas below.

The starting material 1-Scheme I may be obtained from the commerciallyavailable 4,6-dihydroxy-2-methylmercaptopyrimidine by known literatureprocedures, such as those noted in Santilli et al., J. Heterocycl. Chem.(1971), 445-53, wherein POCl₃ and DMF are used.

The intermediate 2-Scheme I was produced by two different routes. In thefirst route, coupling of dichloro aldehyde 1-Scheme I with aryl aminesin the presence of NaH in DMSO (Santilli et al., J. Heterocycl. Chem.(1971), 445-53) afforded the desired compound 2-Scheme I along withimine 13-Scheme I. The imine was converted to aldehyde 2-Scheme I bytreatment with aqueous HCl in THF. Conversion of 1-Scheme 1 to 2-SchemeI may also be achieved using triethylamine and the desired amine inchloroform at room temperature for 10 minutes. The reaction was veryeffective for a range of alkyl amines (78-95% yield). For aryl amines,elevated temperatures (reflux) and longer reaction time (24 hours) werenecessary for reaction completion. Use of the base could be omitted when3 or more equivalent of amine were used. Other suitable bases, includebut are not limited to pyridine, diisopropyl ethylamine or pyrrolidine,which may also be used in an appropriate organic solvent, including butnot limited to THF, diethyl ether or dioxane.

In the second route, the nitrile 9-Scheme I was prepared in three stepsfrom the aldehyde 1-Scheme I (Santilli et al., J. Heterocycl. Chem.(1971), 445-53). Coupling of dichloro nitrile 9-Scheme I with arylamines in the presence of NaH in DMSO afforded the desired compound10-Scheme I. Other suitable bases such as pyridine, diisopropylethylamine, or sodium may also be used in an appropriate organic solventsuch as THF, DMF or dioxane. Production and use of the nitrile9-Scheme-I may also be found in PCT/US01/06688, filed Mar. 2, 2001,published as WO 01/64679 whose disclosure is incorporated herein byreference in its entirety.

The nitrile 10-Scheme I was easily reduced with DIBAL in dichloromethaneat room temperature Boschelliat et al., J. Med. Chem. (1998), 4365-4377)to afford desired 2-Scheme I along with the unsubstituted imine13-Scheme I (R═H). The latter was hydrolyzed to 2-Scheme I in situ withHCl. Other reduction agents, such as lithium aluminum hydride, Raney Ni,or SnCl₂, may be utilized in an appropriate organic solvent such as THE,diethyl ether or dioxane to perform the conversion of 10-Scheme 1 to2-Scheme I.

Aldehyde 2-Scheme I was coupled to arylboronic acids under Suzukicoupling conditions, using a palladium catalyst, such astetrakis(triphenylphosphine) palladium(0), to afford good to excellentyields of 3-Scheme I. Alternatively, the bi-aryl coupling reaction of2-Scheme I may be performed using aryl or heteroaryl organozinc,organocopper, organotin, or other organometallic reagents known toafford bi-aryl cross-coupling products such as 3-Scheme I [see forexample Solberg, J.; Undheim, K. Acta Chemica Scandinavia 1989, 62-68].Displacement of the chlorine in 2-Scheme I may also be achieved withnitrogen nucleophiles [for related aminations see U.S. Pat. Nos.3,631,045 and 3,910,913], sulphur nucleophiles, [see Tumkevicius, S.Liebigs Ann. 1995, 1703-1705], oxygen nucleophiles, or alkylnucleophiles.

3-Scheme I was then converted to pyridopyrimidinone 5-Scheme I by one ofthree procedures. The first procedure used the Wittig reaction, asmodified by Horner-Emmons, converting 3-Scheme 1 to 4-Scheme I. In thisreaction, the aldehyde 3-Scheme I was treated with a suitable phosphorusylide, such as triethyl phosphonoacetate or methyldiethylphosphonoacetate, to give the olefin intermediate 4-Scheme I. Thereaction was performed under reflux, in a suitable base, such as sodiumhydride, sodium methoxide, or sodium hydroxide, and in a suitableorganic solvent such as diethyl ether, dioxane or ethanol. Theconversion of 3-Scheme 1 to 4-Scheme I may also be performed using thePeterson olefination reaction, or an aldol-based olefination reactionthat utilizes acetic anhydride, malonic acid and its monoalkyl esters,or ethyl acetate.

Heating of 4-Scheme I in toluene at 220° C. in a sealed tube (Matyus etal. Heterocycles (1985), 2057-64), followed by solvent removal, affordedthe desired product 5-Scheme I. This reaction may be run in the presenceof a suitable base, such as DBU or diisopropylethyl amine, pyridine,lithium bi(trimethylsilyl)amide, or LDA and in an appropriate organicsolvent such as an organic hydrocarbon, cresol, dioxane, DMF, pyridine,or xylene.

The second procedure used a Horner-Emmons reaction with Stillmodification (Still et al., Tetrahedron Lett. (1983), 4405-8; Jacobsenet al., Tetrahedron (1994), 4323-34) to produce a mixture of desiredproduct 5-Scheme I and trans isomer 4-Scheme I. Trans isomer 4-Scheme Iwas isolated and converted to the desired product 5-Scheme I by heatingto 220° C. in toluene in a sealed tube as described above.

The third procedure involved acetylation of 3-Scheme I, followed by theintramolecular aldol condensation, promoted by an acetylating agent(such as acetic anhydride, acetyl chloride, or a ketene) and a suitablebase (such as pyridine, diispropyl ethylamine, or pyrrolidine), togenerate 5-Scheme I in a very good yield. The third procedure is optimalwhen R₃ is an optionally substituted aryl, or heteroaryl. When R₃ is anarylalkyl, or heteroarylalkyl substituent it is not clear that thereaction will form the key intermediate of Formula (VII), as shown below(3a-Scheme II), which may optionally be isolated, as shown in Scheme IIbelow. Compounds of Formula (VI) are preferably not isolated but furtherreacted with a base or with heat to cyclize into 5-Scheme-I. The firstand second procedures should be utilized for all other R₃ moieties.

Oxidation of the sulfide 5-Scheme I to the sulfone 6-Scheme I wasperformed using meta-chloroperoxybenzoic acid (mCPBA) in high yield andpurity. Suitable oxidation methods for use herein include use of one ortwo equivalents of meta-chloroperoxybenzoic acid (mCPBA) or Oxone® toafford either the sulfoxides or sulfones. Oxidation of the sulfides tosulfoxides or sulfones can also be effected by OsO₄ and catalytictertiary amine N-oxide, hydrogen peroxide, other peracids, oxygen,ozone, organic peroxides, potassium and zinc permanganate, potassiumpersulfate, and sodium hypochlorite.

Displacements of the sulfones 6-Scheme I to the final products7-Scheme-I were usually done with an excess of amine inN-methylpyrrolidine (Barvian et al., J. Med. Chem. (2000), 4606-4616). Awide range of primary amines underwent this reaction with excellentyields. In some cases (in O-displacement or sulfonamide formation) ananion of the nucleophile was prepared with base (usually sodium hydride)in dimethylformamide and then added to the sulfone. Yields for thesereactions were usually lower. Similarly related sulfones and sulfoxidesof the compounds herein wherein X is SO-alkyl or SO₂-alkyl have beenreported in the literature to be displaced by a wide variety ofnucleophiles. Thus the analogs of the compounds herein wherein X is analkyl sulfone or sulfoxide may be displaced by primary and secondaryalkylamines without additional base catalysis, preferably in a polaraprotic solvent, such as but not limited to, N-methylpyrrolidin-2-one(NMP), and at varying temperatures depending upon the nucleophilicity ofthe amine. For instance displacement of the sulfone of analogs ofFormula (I) compounds with ethanolamine, in NMP, occurred in 30 min. at65° C., while a more hindered amine such astris(hydroxymethyl)-aminomethane may require elevated temperatures andextended reaction times (80° C. over a 24 hour reaction time). Thesulfone may also be displaced with a substituted arylamine, orheteroarylamine at elevated temperatures, sometimes requiring formationof the aryl or heteroarylamine anion with sodium hydride, or othersuitable base, in DMSO. In addition, the sulfoxide analogs of Formula(I) compounds may be readily displaced with aluminum salts of aryl orheteroaryl amines as previously described in the patent literature (WO99/32121). Likewise, sulfone and sulfoxide analogs of Formula (I) and(Ia) may be displaced with aryl or heteroaryl or alkyl thiols or alkylor aryl or heteroaryl alcohols. For instance analogs of (I) containingsulfones as the X substituents may be displaced with sodium alkoxide inthe alcohol, or alternatively reactive alkoxide or phenoxidenucleophiles may be generated from the alcohol or phenol with a suitablebase such as sodium, NaH or sodium bistrimethylsilyl amide in a polaraprotic solvent such as DMSO, or run as a neat reaction. Similarlysulfones related to Formula (I) and (Ia), for instance, may be displacedwith carbon nucleophiles such as aryl or alkyl Grignard reagents orrelated organometallics such as organo lithium, zinc, tin or boron.These reactions may, in some cases, require transition metal catalysissuch as with Pd or Ni catalysts. Displacement of related 2-pyrimidinesulfones with cyanide, malonate anions, unactivated enolates, orheterocyclic C nucleophiles such as 1-methylimidazole anion, by thegeneration of the anion with NaH or other suitable base in THF also hasprecedent (see for example, Chem Pharm Bull. 1987, 4972-4976.). Forexample, analogs of Formula (I) and (Ia) compounds wherein X is an alkylsulfone may be displaced with the anion of 1-methyl imidazole, generatedby treatment of 1-methyl imidazole with n-butyl lithium in a solventsuch as THF at temperatures of about −70°, to afford the C-alkylatedproduct substituted on the imidazole C-2.

For the purposes herein, compounds of Formulas (I), (Ia), (II) and (IIa)wherein X is R₂ or NHS(O)mR₂ may be obtained from compounds of 6-SchemeI by displacement of the sulfone using the appropriate “X” functionalityas defined in Formula (I) and (Ia). To obtain compounds of Formulas (I),(Ia), (II) and (IIa) wherein X is S(O)_(m)R₂ and R₂ is other thanmethyl, displacement of the sulfone on the corresponding compound6-Scheme I by thiol (R₂SH) and then followed by oxidation, if desired,with an appropriate oxidating agent, such as MCPBA, or KMnO₄. Suitableoxidation methods for use herein include use of an oxidant such as oneor two equivalents of meta-chloroperoxybenzoic acid or Oxone® to affordeither the sulfoxides or sulfones. Oxidation of the sulfides to sulfonesmay also be effected by OSO₄ and catalytic tertiary amine N-oxide. Othermethods for sulfide oxidation include the use of hydrogen peroxide,other peracids, oxygen, ozone, organic peroxides, potassium and zincpermanganate, potassium persulfate, and sodium hypochlorite.

8-Scheme I can be also prepared by heating the trans ester 4-Scheme I inalcohol in the presence of the corresponding sodium alkoxide. The yieldof this reaction was very high for primary alcohols, but longer reactiontimes were required for secondary alcohols. Sodium alkoxides may beeasily prepared from corresponding alcohol and base, such as sodium orsodium hydride.

Reduction of trans ester 4-Scheme I with SmI₂ gives the reduced analogue11-Scheme I. This reduction can be also done in the presence of otherreducing agents such as hydrogen gas, lithium in liquid ammonia,magnesium or sodium borohydride in the appropriate organic solvent suchas THF, ethanol or diethyl ether.

Cyclization of the ester 11-Scheme I can be done utilizing sodiummethoxide in methanol to give reduced analogue 12-Scheme I. Otherorganic bases, such as sodium, sodium ethoxide or TEA can be used in anappropriate organic solvent such as methanol, ethanol or dioxane. Theproduct 12-Scheme I can be also obtained by heating ester 11-Scheme I to150° C. in an appropriate organic solvent, such as toluene, xylene orisopropanol.

Additional procedures for producing similar intermediates to thoseherein, which the skilled artisan may find may be found in WO 99/41253,now U.S. Pat. No. 6,200,977; U.S. Pat. No. 6,153,619; U.S. Pat. No.6,268,310; U.S. Pat. No. 5,468,751; U.S. Pat. No. 5,474,996; and EP 1040 831.

An illustration of an alternative preparation of compounds of Formula(VII) the present invention is shown in Scheme II below, and describedabove.

Another aspect of the present invention are novel intermediates of theformula (III)

wherein

-   R₁ is an aryl or heteroaryl ring, which ring is optionally    substituted;-   R₃ is an C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, C₃₋₇ cycloalkylalkyl, aryl,    arylC₁₋₁₀ alkyl, heteroaryl, heteroarylC₁₋₁₀ alkyl, heterocyclic, or    a heterocyclylC₁₋₁₀ alkyl moiety, which moieties are optionally    substituted;-   R₁₂ is a C₁₋₁₀ alkyl, aryl, heteroaryl, or arylalkyl;-   m is 0 or an integer having a value of 1 or 2; and-   Rg is a C₁₋₄ alkyl.

Preferably, Rg is a C₁₋₄ alkyl, and more preferably methyl.

Preferably, m is 0 or an integer having a value of 1 or 2. Morepreferably m is 0 or 2.

Preferably, R₁ is an aryl moiety, more preferably a phenyl ring,optionally substituted one or more times by halogen, C₁₋₄ alkyl, orhalo-substituted-C₁₋₄ alkyl. More preferably, the phenyl ring issubstituted independently in the 2, 4, or 6-positions, or di-substitutedin the 2,4- positions, such as 2-fluoro, 4-fluoro, 2,4-difluoro,2,4,6-trifluoro, or 2-methyl-4-fluoro. More preferably, the phenyl ringis independently substituted in the 3 position, 2,3-disubstituted or3,4-disubstituted, such as by fluorine or chlorine.

Another aspect of the present invention are novel intermediates of theformula (IIIa)

wherein

-   R₁ is the moiety YRa;-   Y is C(R_(b))(R_(d)), C(O), N(R_(d)), N(R_(d))C(R_(c))(R_(d)),    oxygen, OC(R_(c))(R_(d)), S(O)m, or S(O)_(m)C(R_(c))(R_(d));-   R_(a) is an aryl or heteroaryl ring, which ring is optionally    substituted;-   R_(b) is hydrogen, C₁₋₂ alkyl, NR_(c), hydroxy, thio, C₁₋₂ alkoxy,    S(O)_(m)C₁₋₂ alkyl;-   R_(c) is hydrogen or C₁₋₂ alkyl;-   R_(d) is hydrogen or C₁₋₂ alkyl;-   m is 0 or an integer having a value of 1 or 2; and-   R₃ is an C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, C₃₋₇ cycloalkylalkyl, aryl,    arylC₁₋₁₀ alkyl, heteroaryl, heteroarylC₁₋₁₀ alkyl, heterocyclic, or    a heterocyclylC₁₋₁₀ alkyl moiety, which moieties are optionally    substituted;-   R₁₂ is a C₁₋₁₀ alkyl, aryl, heteroaryl, or arylalkyl;-   m is 0 or an integer having a value of 1 or 2; and-   Rg is a C₁₋₄ alkyl.

The substituents of compounds of Formula (II) and (IIa), and (IV) and(IVa) below follow those preferances of the final compounds of Formula(I) or (II) herein, respectively.

Another aspect of the present invention are the novel intermediates ofthe formula (IV)

wherein R₁, R₃, R₁₂, m and R_(g) are as defined for Formula (III) above.

Another aspect of the present invention are the novel intermediates ofthe formula (IVa)

wherein R₁, R₃, R₁₂, m and R_(g) are as defined for Formula (IIIa)above.

Another aspect of the present invention are novel intermediates of theformula (IV)

wherein R₁, R₃, R₁₂, m and R_(g) are as defined for Formula (III) above.

Another aspect of the present invention are novel intermediates of theformula (IVa)

wherein R₁, R₃, R₁₂, Rg and m are as defined for Formula (IIIa) above.

Another aspect of the present invention are novel intermediates of theformula

wherein

-   R₁ is a halogen, an optionally substituted aryl or an optionally    substituted heteroaryl ring;-   R₃ is hydrogen, C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, C₃₋₇ cycloalkylalkyl,    aryl, arylC₁₋₁₀ alkyl, heteroaryl, heteroarylC₁₋₁₀ alkyl,    heterocyclic, or a heterocyclylC₁₋₁₀ alkyl moiety, which moieties    are optionally substituted; provided that when R₃ is hydrogen, then    R₁ is other than chlorine;-   m is 0 or an integer having a value of 1 or 2; and-   Rg is a C₁₋₄ alkyl

Preferably, R₁ is a halogen, more preferably chlorine, or an arylmoiety, more preferably a phenyl ring, optionally substituted one ormore times independently by halogen, C₁₋₄ alkyl, orhalo-substituted-C₁₋₄ alkyl. More preferably, the phenyl ring issubstituted in the 2, 4, or 6-positions, or di-substituted in the 2,4-positions, such as 2-fluoro, 4-fluoro, 2,4-difluoro, 2,4,6-trifluoro, or2-methyl-4-fluoro. Alternatively, the phenyl ring is independentlysubstituted in the 3 position, 2,3-disubstituted or 3,4-disubstituted,such as by fluorine or chlorine.

Preferably, R₃ is an optionally substituted C₁₋₁₀ alkyl, C₃₋₇cycloalkyl, C₃₋₇ cycloalkylalkyl, or aryl.

Preferably, the R₃ optional substituents are independently selected fromC₁₋₁₀ alkyl, halo-substituted C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl,C₃₋₇ cycloalkyl, C₃₋₇cycloalkylC₁₋₁₀ alkyl, C₅₋₇ cycloalkenyl, C₅₋₇cycloalkenyl C₁₋₁₀ alkyl, halogen, (CR₁₀R₂₀)_(n)OR₆, (CR₁₀R₂₀)_(n)SH,(CR₁₀R₂₀)_(n)S(O)_(m)R₇, (CR₁₀R₂₀)_(n)NHS(O)₂R₇, (CR₁₀R₂₀)_(n)NR₄R₁₄,(CR₁₀R₂₀)_(n)CN, (CR₁₀R₂₀)_(n) S(O)₂NR₄R₁₄, (CR₁₀R₂₀)_(n)C(Z)R₆,(CR₁₀R₂₀)_(n)OC(Z)R₆, (CR₁₀R₂₀)_(n)C(Z)OR₆, (CR₁₀R₂₀)_(n)C(Z)NR₄R₁₄,(CR₁₀R₂₀)_(n)NR₁₀C(Z)R₆, (CR₁₀R₂₀)_(n)NR₁₀C(═NR₁₀) NR₄R₁₄,(CR₁₀R₂₀)_(n)OC(Z)NR₄R₁₄, (CR₁₀R₂₀)_(n)NR₁₀C(Z) NR₄R₁₄, or(CR₁₀R₂₀)_(n)NR₁₀C(Z)OR₇.

More preferably, the optional substituents are independently selectedfrom halogen, alkyl, hydroxy, alkoxy, amino, or halosubstituted alkyl.

Exemplified compounds of Formula (VI) include, but are not limited to:

-   4-(2,4-difluoro-phenylamino)-6-(2,4,6-trifluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde;-   4-(2,4-difluoro-phenyl)-6-(2,4-difluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde;-   4-(2-fluorophenyl)-6-(2,4,6-trifluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde;-   4-(4-fluorophenyl)-6-(2,4,6-trifluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde;-   4-(2-fluorophenyl)-6-(2,4-difluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde;-   4-(2,4-Difluoro-phenyl)-6-(4-fluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde;-   4-(2,4-difluoro-phenyl)-6-(2,4-difluorophenylamino)-methylsulfanyl-pyrimidine-5-carbaldehyde;-   4-(2-chloro-4-fluorophenyl)-6-(2,4-difluorophenyl    amino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde; or-   4-(2,4-difluorophenyl)-6-(2,3-difluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde.

Another aspect of the present invention are novel intermediates of theformula

wherein

-   R₁ is YRa;-   Y is C(R_(b))(R_(d)), C(O), N(R_(d)), N(R_(d))C(R_(c))(R_(d)),    oxygen, OC(R_(c))(R_(d)), S(O)m, or S(O)_(m)C(R_(c))(R_(d));-   R_(a) is an aryl or heteroaryl ring, which ring is optionally    substituted;-   R_(b) is hydrogen, C₁₋₂ alkyl, NR_(C), hydroxy, thio, C₁₋₂ alkoxy,    S(O)_(m)C₁₋₂ alkyl;-   R_(c) is hydrogen or C₁₋₂ alkyl;-   R_(d) is hydrogen or C₁₋₂ alkyl;-   m is 0 or an integer having a value of 1 or 2; and-   R₃ is hydrogen, C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, C₃₋₇ cycloalkylalkyl,    aryl, arylC₁₋₁₀ alkyl, heteroaryl, heteroarylC₁₋₁₀ alkyl,    heterocyclic, or a heterocyclylC₁₋₁₀ alkyl moiety, which moieties    are optionally substituted;-   m is 0 or an integer having a value of 1 or 2; and-   Rg is a C₁₋₄ alkyl.

Preferably, as noted above, the substituents of compounds of Formula(VI) and (VIa) follow those of the final compounds of Formula (I), and(II) herein.

Exemplified compounds of Formula (VI) include, but are not limited to,4-(2-Chloro-phenylamino)-2-methylsulfanyl-6-phenoxy-pyrimidine-5-carbaldehyde.

Another aspect of this invention are novel intermediates of Formula(VII)

wherein

R₁ is as defined above for Formula (I) compounds, and R₃, Rg, and m isan optionally substituted aryl or heteroaryl moiety, as defined forFormula (III) compounds.

Another aspect of this invention are novel intermediates of Formula(VIIa)

wherein

R₁ is defined above for Formula (II) compounds, and R₃, Rg, and m is anoptionally substituted aryl or heteroaryl moiety, as defined for Formula(IIIa) compounds.

Another aspect of the present invention are novel intermediates of theformula

wherein

-   R₁ is a halogen;-   R₃ is hydrogen, C₁₋₁₀ alkyl, C₃₋₇ cycloalkyl, C₃₋₇ cycloalkylalkyl,    aryl, arylC₁₋₁₀ alkyl, heteroaryl, heteroarylC₁₋₁₀ alkyl,    heterocyclic, or a heterocyclylC₁₋₁₀ alkyl moiety, which moieties    are optionally substituted; provided that when R₃ is hydrogen, then    R₁ is other than chlorine;-   m is 0 or an integer having a value of 1 or 2; and-   Rg is a C₁₋₄ alkyl.

Preferably R₁ is a halogen, more preferably chlorine.

Suitably, the R₃ substituents are the same as those for compounds ofFormulas (I) and (II) herein.

Representative compounds of Formula (I) and (Ia) are:

-   8-(2,4,6-trifluorophenyl)-4-(2,4-difluorophenyl)-2-(2-hydroxy-1-hydroxymethylethylamino)-8H-pyridio[2,3-d]pyrimidin-7-one;-   4,8-Bis-(2,4-difluoro-phenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido-[2,3-d]-pyrimidin-7-one;-   8-(2,4,6-trifluorophenyl)-4-(2-fluorophenyl)-2-(2-hydroxy-1-hydroxymethylethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one;-   8-(2,4,6-trifluorophenyl)-4-(4-fluorophenyl)-2-(2-hydroxy-1-hydroxymethylethylamino)-8H-pyridio[2,3-d]pyrimidin-7-one;-   4-(2-fluoro-phenyl)-8-(2,4-difluoro-phenyl)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8H-pyrido-[2,3-d]-pyrimidin-7-one;-   4-(2,4-Difluoro-phenyl)-8-(4-fluoro-phenyl)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8H-pyrido-[2,3-d]-pyrimidin-7-one;-   4,8-Bis-(2,4-difluoro-phenyl)-2-(2-hydroxy-1-methyl-ethylamino)-8H-pyrido-[2,3-d]-pyrimidin-7-one;-   8-(2,4-Difluorophenyl)-4-(2-chloro-4-fluorophenyl)-2-(2-hydroxy-1-hydroxymethylethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one;    or-   8-(2,3-difluorophenyl)-4-(2,4-difluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-    one; or a pharmaceutically acceptable salt thereof.    Methods of Treatment

The compounds of Formula (I) and (Ia) or a pharmaceutically acceptablesalt thereof can be used in the manufacture of a medicament for theprophylactic or therapeutic treatment of any disease state in a human,or other mammal, which is exacerbated or caused by excessive orunregulated cytokine production by such mammal's cell, such as but notlimited to monocytes and/or macrophages.

For purposes herein, compounds of Formula (I) and (Ia) will all bereferred to as compounds of Formula (I) unless otherwise indicated.

Compounds of Formula (I) are capable of inhibiting proinflammatorycytokines, such as IL-1, IL-6, IL-8, and TNF and are therefore of use intherapy. IL-1, IL-6, IL-8 and TNF affect a wide variety of cells andtissues and these cytokines, as well as other leukocyte-derivedcytokines, are important and critical inflammatory mediators of a widevariety of disease states and conditions. The inhibition of thesepro-inflammatory cytokines is of benefit in controlling, reducing andalleviating many of these disease states.

Accordingly, the present invention provides a method of treating acytokine-mediated disease which comprises administering an effectivecytokine-interfering amount of a compound of Formula (I) or apharmaceutically acceptable salt thereof.

Compounds of Formula (I) are capable of inhibiting inducibleproinflammatory proteins, such as COX-2, also referred to by many othernames such as prostaglandin endoperoxide synthase-2 (PGHS-2) and aretherefore of use in therapy. These proinflammatory lipid mediators ofthe cyclooxygenase (CO) pathway are produced by the inducible COX-2enzyme. Regulation, therefore of COX-2 which is responsible for thethese products derived from arachidonic acid, such as prostaglandinsaffect a wide variety of cells and tissues are important and criticalinflammatory mediators of a wide variety of disease states andconditions. Expression of COX-1 is not effected by compounds of Formula(I). This selective inhibition of COX-2 may alleviate or spareulcerogenic liability associated with inhibition of COX-1 therebyinhibiting prostoglandins essential for cytoprotective effects. Thusinhibition of these pro-inflammatory mediators is of benefit incontrolling, reducing and alleviating many of these disease states. Mostnotably these inflammatory mediators, in particular prostaglandins, havebeen implicated in pain, such as in the sensitization of pain receptors,or edema. This aspect of pain management therefore includes treatment ofneuromuscular pain, headache, cancer pain, and arthritis pain. Compoundsof Formula (I) or a pharmaceutically acceptable salt thereof, are of usein the prophylaxis or therapy in a human, or other mammal, by inhibitionof the synthesis of the COX-2 enzyme.

Accordingly, the present invention provides a method of inhibiting thesynthesis of COX-2 which comprises administering an effective amount ofa compound of Formula (I) or a pharmaceutically acceptable salt thereof.The present invention also provides for a method of prophylaxistreatment in a human, or other mammal, by inhibition of the synthesis ofthe COX-2 enzyme.

In particular, compounds of Formula (I) or a pharmaceutically acceptablesalt thereof are of use in the prophylaxis or therapy of any diseasestate in a human, or other mammal, which is exacerbated by or caused byexcessive or unregulated IL-1, IL-6, IL-8 or TNF production by suchmammal's cell, such as, but not limited to, monocytes and/ormacrophages.

Accordingly, in another aspect, this invention relates to a method ofinhibiting the production of IL-1 in a mammal in need thereof whichcomprises administering to said mammal an effective amount of a compoundof Formula (I) or a pharmaceutically acceptable salt thereof.

There are many disease states in which excessive or unregulated IL-1production is implicated in exacerbating and/or causing the disease.These include rheumatoid arthritis, osteoarthritis, meningitis, ischemicand hemorrhagic stroke, neurotrauma/closed head injury, stroke,endotoxemia and/or toxic shock syndrome, other acute or chronicinflammatory disease states such as the inflammatory reaction induced byendotoxin or inflammatory bowel disease, tuberculosis, atherosclerosis,muscle degeneration, multiple sclerosis, cachexia, bone resorption,psoriatic arthritis, Reiter's syndrome, gout, traumatic arthritis,rubella arthritis and acute synovitis.

Recent evidence also links IL-1 activity to diabetes, pancreatic B celldiseases and Alzheimer's disease.

Use of a CSAID inhibitor compound for the treatment of CSBP mediateddisease states, can include, but not be limited to neurodegenerativediseases, such as Alzheimer's disease (as noted above), Parkinson'sdisease and multiple sclerosis, etc.

In a further aspect, this invention relates to a method of inhibitingthe production of TNF in a mammal in need thereof which comprisesadministering to said mammal an effective amount of a compound ofFormula (I) or a pharmaceutically acceptable salt thereof.

Excessive or unregulated TNF production has been implicated in mediatingor exacerbating a number of diseases including rheumatoid arthritis,rheumatoid spondylitis, osteoarthritis, gouty arthritis and otherarthritic conditions, sepsis, septic shock, endotoxic shock, gramnegative sepsis, toxic shock syndrome, adult respiratory distresssyndrome, chronic pulmonary inflammatory disease and chronic obstructivepulmonary disease, silicosis, pulmonary sarcoisosis, bone resorptiondiseases, such as osteoporosis, cardiac, brain and renal reperfusioninjury, graft vs. host reaction, allograft rejections, fever andmyalgias due to infection, such as influenza, brain infections includingencephalitis (including HIV-induced forms), cerebral malaria,meningitis, ischemic and hemorrhagic stroke, cachexia secondary toinfection or malignancy, cachexia secondary to acquired immunedeficiency syndrome (AIDS), AIDS, ARC (AIDS related complex), keloidformation, scar tissue formation, inflammatory bowel disease, Crohn'sdisease, ulcerative colitis and pyresis.

Compounds of Formula (I) are also useful in the treatment of viralinfections, where such viruses are sensitive to upregulation by TNF orwill elicit TNF production in vivo. The viruses contemplated fortreatment herein are those that produce TNF as a result of infection, orthose which are sensitive to inhibition, such as by decreasedreplication, directly or indirectly, by the TNF inhibiting-compounds ofFormula (I). Such viruses include, but are not limited to HIV-1, HIV-2and HIV-3, Cytomegalovirus (CMV), Influenza, adenovirus and the Herpesgroup of viruses, such as but not limited to, Herpes Zoster and HerpesSimplex. Accordingly, in a further aspect, this invention relates to amethod of treating a mammal afflicted with a human immunodeficiencyvirus (HIV) which comprises administering to such mammal an effectiveTNF inhibiting amount of a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof.

It is also recognized that both IL-6 and IL-8 are produced duringrhinovirus (HRV) infections and contribute to the pathogenesis of commoncold and exacerbation of asthma associated with HRV infection (Turner etal. (1998), Clin. Infec. Dis., Vol 26, p 840; Teren et al. (1997), Am JRespir Crit Care Med vol 155, p1362; Grunberg et al. (1997), Am J RespirCrit Care Med 156:609 and Zhu et al, J Clin Invest (1996), 97:421). Ithas also been demonstrated in vitro that infection of pulmonaryepithelial cells with HRV results in production of IL-6 and IL-8(Subauste et al., J. Clin. Invest. 1995, 96:549.) Epithelial cellsrepresent the primary site of infection of HRV. Therefore another aspectof the present invention is a method of treatment to reduce inflammationassociated with a rhinovirus infection, not necessarily a direct effecton virus itself.

Compounds of Formula (I) may also be used in association with theveterinary treatment of mammals, other than in humans, in need ofinhibition of TNF production. TNF mediated diseases for treatment,therapeutically or prophylactically, in animals include disease statessuch as those noted above, but in particular viral infections. Examplesof such viruses include, but are not limited to, lentivirus infectionssuch as, equine infectious anaemia virus, caprine arthritis virus, visnavirus, or maedi virus or retrovirus infections, such as but not limitedto feline immunodeficiency virus (FIV), bovine immunodeficiency virus,or canine immunodeficiency virus or other retroviral infections.

The compounds of Formula (I) may also be used topically in the treatmentor prophylaxis of topical disease states mediated by or exacerbated byexcessive cytokine production, such as by IL-1 or TNF respectively, suchas inflamed joints, eczema, psoriasis and other inflammatory skinconditions such as sunburn; inflammatory eye conditions includingconjunctivitis; pyrexia, pain and other conditions associated withinflammation. Periodontal disease has also been implemented in cytokineproduction, both topically and systemically. Hence use of compounds ofFormula (I) to control the inflammation associated with cytokineproduction in such peroral diseases such as gingivitis and periodontitisis another aspect of the present invention.

Compounds of Formula (I) have also been shown to inhibit the productionof IL-8 (Interleukin-8, NAP). Accordingly, in a further aspect, thisinvention relates to a method of inhibiting the production of IL-8 in amammal in need thereof which comprises administering to said mammal aneffective amount of a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof.

There are many disease states in which excessive or unregulated IL-8production is implicated in exacerbating and/or causing the disease.These diseases are characterized by massive neutrophil infiltration suchas, psoriasis, inflammatory bowel disease, asthma, cardiac, brain andrenal reperfusion injury, adult respiratory distress syndrome,thrombosis and glomerulonephritis. All of these diseases are associatedwith increased IL-8 production which is responsible for the chemotaxisof neutrophils into the inflammatory site. In contrast to otherinflammatory cytokines (IL-1, TNF, and IL-6), IL-8 has the uniqueproperty of promoting neutrophil chemotaxis and activation. Therefore,the inhibition of IL-8 production would lead to a direct reduction inthe neutrophil infiltration.

The compounds of Formula (I) are administered in an amount sufficient toinhibit cytokine, in particular IL-1, IL-6, IL-8 or TNF, production suchthat it is regulated down to normal levels, or in some case to subnormallevels, so as to ameliorate or prevent the disease state. Abnormallevels of IL-1, IL-6, IL-8 or TNF, for instance in the context of thepresent invention, constitute: (i) levels of free (not cell bound) IL-1,IL-6, IL-8 or TNF greater than or equal to 1 picogram per ml; (ii) anycell associated IL-1, IL-6, IL-8 or TNF; or (iii) the presence of IL-1,IL-6, IL-8 or TNF mRNA above basal levels in cells or tissues in whichIL-1, IL-6, IL-8 or TNF, respectively, is produced.

The discovery that the compounds of Formula (I) are inhibitors ofcytokines, specifically IL-1, IL-6, EL-8 and TNF is based upon theeffects of the compounds of Formulas (I) on the production of the IL-1,IL-8 and TNF in in vitro assays which are described herein.

As used herein, the term “inhibiting the production of IL-1 (IL-6, IL-8or TNF)” refers to:

a) a decrease of excessive in vivo levels of the cytokine (IL-1, IL-6,IL-8 or TNF) in a human to normal or sub-normal levels by inhibition ofthe in release of the cytokine by all cells, including but not limitedto monocytes or macrophages;

b) a down regulation, at the genomic level, of excessive in vivo levelsof the cytokine (IL-1, IL-6, IL-8 or TNF) in a human to normal orsub-normal levels;

c) a down regulation, by inhibition of the direct synthesis of thecytokine (IL-1, IL-6, IL-8 or TNF) as a postranslational event; or

d) a down regulation, at the translational level, of excessive in vivolevels of the cytokine (IL-1, IL-6, IL-8 or TNF) in a human to normal orsub-normal levels.

As used herein, the term “TNF mediated disease or disease state” refersto any and all disease states in which TNF plays a role, either byproduction of TNF itself, or by TNF causing another monokine to bereleased, such as but not limited to IL-1, IL-6 or IL-8. A disease statein which, for instance, IL-1 is a major component, and whose productionor action, is exacerbated or secreted in response to TNF, wouldtherefore be considered a disease stated mediated by TNF.

As used herein, the term “cytokine” refers to any secreted polypeptidethat affects the functions of cells and is a molecule which modulatesinteractions between cells in the immune, inflammatory or hematopoieticresponse. A cytokine includes, but is not limited to, monokines andlymphokines, regardless of which cells produce them. For instance, amonokine is generally referred to as being produced and secreted by amononuclear cell, such as a macrophage and/or monocyte. Many other cellshowever also produce monokines, such as natural killer cells,fibroblasts, basophils, neutrophils, endothelial cells, brainastrocytes, bone marrow stromal cells, epideral keratinocytes andB-lymphocytes. Lymphokines are generally referred to as being producedby lymphocyte cells. Examples of cytokines include, but are not limitedto, Interleukin-1 (IL-1), Interleukin-6 (IL-6), Interleukin-8 (IL-8),Tumor Necrosis Factor-alpha (TNF-a) and Tumor Necrosis Factor beta(TNF-β).

As used herein, the term “cytokine interfering” or “cytokine suppressiveamount” refers to an effective amount of a compound of Formula (I) whichwill cause a decrease in the in vivo levels of the cytokine to normal orsub-normal levels, when given to a patient for the prophylaxis ortreatment of a disease state which is exacerbated by, or caused by,excessive or unregulated cytokine production.

As used herein, the cytokine referred to in the phrase “inhibition of acytokine, for use in the treatment of a HIV-infected human” is acytokine which is implicated in (a) the initiation and/or maintenance ofT cell activation and/or activated T cell-mediated HIV gene expressionand/or replication and/or (b) any cytokine-mediated disease associatedproblem such as cachexia or muscle degeneration.

As TNF-β (also known as lymphotoxin) has close structural homology withTNF-α (also known as cachectin) and since each induces similar biologicresponses and binds to the same cellular receptor, both TNF-α and TNF-βare inhibited by the compounds of the present invention and thus areherein referred to collectively as “TNF” unless specifically delineatedotherwise.

A member of the MAP kinase family, alternatively termed CSBP, p38, orRK, has been identified independently by several laboratories.Activation of this novel protein kinase via dual phosphorylation hasbeen observed in different cell systems upon stimulation by a widespectrum of stimuli, such as physicochemical stress and treatment withlipopolysaccharide or proinflammatory cytokines such as interleukin-1and tumor necrosis factor. The cytokine biosynthesis inhibitors, of thepresent invention, compounds of Formula (I) have been determined to bepotent and selective inhibitors of CSBP/p38/RK kinase activity. Theseinhibitors are of aid in determining the signaling pathways involvementin inflammatory responses. In particular, for the first time adefinitive signal transduction pathway can be prescribed to the actionof lipopolysaccharide in cytokine production in macrophages. In additionto those diseases already noted, treatment of stroke, neurotrauma,cardiac and renal reperfusion injury, congestive heart failure, coronaryarterial bypass grafting (CABG) surgery, chronic renal failure,angiogenesis & related processes, such as cancer, thrombosis,glomerulonephritis, diabetes and pancreatic P cells, multiple sclerosis,muscle degeneration, eczema, psoriasis, sunburn, and conjunctivitis arealso included.

The CSBP inhibitors were subsequently tested in a number of animalmodels for anti-inflammatory activity. Model systems were chosen thatwere relatively insensitive to cyclooxygenase inhibitors in order toreveal the unique activities of cytokine suppressive agents. Theinhibitors exhibited significant activity in many such in vivo studies.Most notable are its effectiveness in the collagen-induced arthritismodel and inhibition of TNF production in the endotoxic shock model. Inthe latter study, the reduction in plasma level of TNF correlated withsurvival and protection from endotoxic shock related mortality. Also ofgreat importance are the compounds effectiveness in inhibiting boneresorption in a rat fetal long bone organ culture system. Griswold etal., (1988) Arthritis Rheum. 31:1406-1412; Badger, et al., (1989) Circ.Shock 27, 51-61; Votta et al., (1994) in vitro. Bone 15, 533-538; Lee etal., (1993). B Ann. N.Y. Acad. Sci. 696, 149-170.

Chronic diseases which have an inappropriate angiogenic component arevarious ocular neovasularizations, such as diabetic retinopathy andmacular degeneration. Other chronic diseases which have an excessive orincreased proliferation of vasculature are tumor growth and metastasis,atherosclerosis, and certain arthritic conditions. Therefore CSBP kinaseinhibitors will be of utility in the blocking of the angiogeniccomponent of these disease states.

The term “excessive or increased proliferation of vasculatureinappropriate angiogenesis” as used herein includes, but is not limitedto, diseases which are characterized by hemangiomas and ocular diseases.

The term “inappropriate angiogenesis” as used herein includes, but isnot limited to, diseases which are characterized by vesicleproliferation with accompanying tissue proliferation, such as occurs incancer, metastasis, arthritis and atherosclerosis.

Accordingly, the present invention provides a method of treating a CSBPkinase mediated disease in a mammal in need thereof, preferably a human,which comprises administering to said mammal, an effective amount of acompound of Formula (I) or a pharmaceutically acceptable salt thereof.

In order to use a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof in therapy, it will normally be formulated intoa pharmaceutical composition in accordance with standard pharmaceuticalpractice. This invention, therefore, also relates to a pharmaceuticalcomposition comprising an effective, non-toxic amount of a compound ofFormula (I) and a pharmaceutically acceptable carrier or diluent.

Compounds of Formula (I), pharmaceutically acceptable salts thereof andpharmaceutical compositions incorporating such may conveniently beadministered by any of the routes conventionally used for drugadministration, for instance, orally, topically, parenterally or byinhalation. The compounds of Formula (I) may be administered inconventional dosage forms prepared by combining a compound of Formula(I) with standard pharmaceutical carriers according to conventionalprocedures. The compounds of Formula (I) may also be administered inconventional dosages in combination with a known, second therapeuticallyactive compound. These procedures may involve mixing, granulating andcompressing or dissolving the ingredients as appropriate to the desiredpreparation. It will be appreciated that the form and character of thepharmaceutically acceptable character or diluent is dictated by theamount of active ingredient with which it is to be combined, the routeof administration and other well-known variables. The carrier(s) must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not deleterious to the recipient thereof.

The pharmaceutical carrier employed may be, for example, either a solidor liquid. Exemplary of solid carriers are lactose, terra alba, sucrose,talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acidand the like. Exemplary of liquid carriers are syrup, peanut oil, oliveoil, water and the like. Similarly, the carrier or diluent may includetime delay material well known to the art, such as glycerylmono-stearate or glyceryl distearate alone or with a wax.

A wide variety of pharmaceutical forms can be employed. Thus, if a solidcarrier is used, the preparation can be tableted, placed in a hardgelatin capsule in powder or pellet form or in the form of a troche orlozenge. The amount of solid carrier will vary widely but preferablywill be from about 25 mg. to about 1 g. When a liquid carrier is used,the preparation will be in the form of a syrup, emulsion, soft gelatincapsule, sterile injectable liquid such as an ampule or nonaqueousliquid suspension.

Compounds of Formula (I) may be administered topically, that is bynon-systemic administration. This includes the application of a compoundof Formula (I) externally to the epidermis or the buccal cavity and theinstillation of such a compound into the ear, eye and nose, such thatthe compound does not significantly enter the blood stream. In contrast,systemic administration refers to oral, intravenous, intraperitoneal andintramuscular administration.

Formulations suitable for topical administration include liquid orsemi-liquid preparations suitable for penetration through the skin tothe site of inflammation such as liniments, lotions, creams, ointmentsor pastes, and drops suitable for administration to the eye, ear ornose. The active ingredient may comprise, for topical administration,from 0.001% to 10% w/w, for instance from 1% to 2% by weight of theformulation. It may however comprise as much as 10% w/w but preferablywill comprise less than 5% w/w, more preferably from 0.1% to 1% w/w ofthe formulation.

Lotions according to the present invention include those suitable forapplication to the skin or eye. An eye lotion may comprise a sterileaqueous solution optionally containing a bactericide and may be preparedby methods similar to those for the preparation of drops. Lotions orliniments for application to the skin may also include an agent tohasten drying and to cool the skin, such as an alcohol or acetone,and/or a moisturizer such as glycerol or an oil such as castor oil orarachis oil.

Creams, ointments or pastes according to the present invention aresemi-solid formulations of the active ingredient for externalapplication. They may be made by mixing the active ingredient infinely-divided or powdered form, alone or in solution or suspension inan aqueous or non-aqueous fluid, with the aid of suitable machinery,with a greasy or non-greasy base. The base may comprise hydrocarbonssuch as hard, soft or liquid paraffin, glycerol, beeswax, a metallicsoap; a mucilage; an oil of natural origin such as almond, corn,arachis, castor or olive oil; wool fat or its derivatives or a fattyacid such as steric or oleic acid together with an alcohol such aspropylene glycol or a macrogel. The formulation may incorporate anysuitable surface active agent such as an anionic, cationic or non-ionicsurfactant such as a sorbitan ester or a polyoxyethylene derivativethereof. Suspending agents such as natural gums, cellulose derivativesor inorganic materials such as silicaceous silicas, and otheringredients such as lanolin, may also be included.

Drops according to the present invention may comprise sterile aqueous oroily solutions or suspensions and may be prepared by dissolving theactive ingredient in a suitable aqueous solution of a bactericidaland/or fungicidal agent and/or any other suitable preservative, andpreferably including a surface active agent. The resulting solution maythen be clarified by filtration, transferred to a suitable containerwhich is then sealed and sterilized by autoclaving or maintaining at98-100° C. for half an hour. Alternatively, the solution may besterilized by filtration and transferred to the container by an aseptictechnique. Examples of bactericidal and fungicidal agents suitable forinclusion in the drops are phenylmercuric nitrate or acetate (0.002%),benzalkonium chloride (0.01%) and chlorhexidine acetate (0.01%).Suitable solvents for the preparation of an oily solution includeglycerol, diluted alcohol and propylene glycol.

Compounds of Formula (I) may be administered parenterally, that is byintravenous, intramuscular, subcutaneous intranasal, intrarectal,intravaginal or intraperitoneal administration. The subcutaneous andintramuscular forms of parenteral administration are generallypreferred. Appropriate dosage forms for such administration may beprepared by conventional techniques. Compounds of Formula (I) may alsobe administered by inhalation, that is by intranasal and oral inhalationadministration. Appropriate dosage forms for such administration, suchas an aerosol formulation or a metered dose inhaler, may be prepared byconventional techniques.

For all methods of use disclosed herein for the compounds of Formula(I), the daily oral dosage regimen will preferably be from about 0.1 toabout 80 mg/kg of total body weight, preferably from about 0.2 to 30mg/kg, more preferably from about 0.5 mg to 15 mg. The daily parenteraldosage regimen about 0.1 to about 80 mg/kg of total body weight,preferably from about 0.2 to about 30 mg/kg, and more preferably fromabout 0.5 mg to 15 mg/kg. The daily topical dosage regimen willpreferably be from 0.1 mg to 150 mg, administered one to four,preferably two or three times daily. The daily inhalation dosage regimenwill preferably be from about 0.01 mg/kg to about 1 mg/kg per day. Itwill also be recognized by one of skill in the art that the optimalquantity and spacing of individual dosages of a compound of Formula (I)or a pharmaceutically acceptable salt thereof will be determined by thenature and extent of the condition being treated, the form, route andsite of administration, and the particular patient being treated, andthat such optimums can be determined by conventional techniques. It willalso be appreciated by one of skill in the art that the optimal courseof treatment, i.e., the number of doses of a compound of Formula (I) ora pharmaceutically acceptable salt thereof given per day for a definednumber of days, can be ascertained by those skilled in the art usingconventional course of treatment determination tests.

The novel compounds of Formula (I) may also be used in association withthe veterinary treatment of mammals, other than humans, in need ofinhibition of CSBP/p38 or cytokine inhibition or production. Inparticular, CSBP/p38 mediated diseases for treatment, therapeutically orprophylactically, in animals include disease states such as those notedherein in the Methods of Treatment section, but in particular viralinfections. Examples of such viruses include, but are not limited to,lentivirus infections such as, equine infectious anaemia virus, caprinearthritis virus, visna virus, or maedi virus or retrovirus infections,such as but not limited to feline immunodeficiency virus (FIV), bovineimmunodeficiency virus, or canine immunodeficiency virus or otherretroviral infections.

Another aspect of the present invention is a method of treating thecommon cold or respiratory viral infection caused by human rhinovirus(HRV), other enteroviruses, coronavirus, influenza virus, parainfluenzavirus, respiratory syncytial virus, or adenovirus in a human in needthereof which method comprises administering to said human an effectiveamount of a CBSP/p38 inhibitor.

Another aspect of the present invention is a method of treating,including prophylaxis of influenza induced pneumonia in a human in needthereof which method comprises administering to said human an effectiveamount of a CBSP/p38 inhibitor

The present invention also relates to the use of the CSBP/p38 kinaseinhibitor for the treatment, including prophylaxis, of inflammationassociated with a viral infection of a human rhinovirus (HRV), otherenteroviruses, coronavirus, influenza virus, parainfluenza virus,respiratory syncytial virus, or adenovirus. In particular, the presentinvention is directed to the treatment of a viral infection in a human,which is caused by the human rhinovirus (HRV), other enterovirus,coronavirus, influenza virus, parainfluenza virus, respiratory syncytialvirus, or an adenovirus. In particular the invention is directed torespiratory viral infections that exacerbate asthma (induced by suchinfections), chronic bronchitis, chronic obstructive pulmonary disease,otitis media, and sinusitis. While inhibiting IL-8 or other cytokinesmay be beneficial in treating a rhinovirus may be known, the use of aninhibitor of the p38 kinase for treating HRV or other respiratory viralinfections causing the common cold is believed novel. It should be notedthat the respiratory viral infection treated herein may also beassociated with a secondary bacterial infection, such as otitis media,sinusitis, or pneumonia.

For use herein treatment may include prophylaxis for use in a treatmentgroup susceptible to such infections. It may also include reducing thesymptoms of, ameliorating the symptoms of, reducing the severity of,reducing the incidence of, or any other change in the condition of thepatient, which improves the therapeutic outcome.

It should be noted that the treatment herein is not directed to theelimination or treatment of the viral organism itself but is directed totreatment of the respiratory viral infection that exacerbates otherdiseases or symptoms of disease, such as asthma (induced by suchinfections), chronic bronchitis, chronic obstructive pulmonary disease,otitis media, and sinusitis.

A preferred virus for treatment herein is the human rhinovirus infection(HRV) or respiratory syncytial virus (RSV).

The invention will now be described by reference to the followingbiological examples which are merely illustrative and are not to beconstrued as a limitation of the scope of the present invention.

BIOLOGICAL EXAMPLES

The cytokine-inhibiting effects of compounds of the present inventionmay be determined by the following in vitro assays: Assays forInterleukin -1 (IL-1), Interleukin -8 (IL-8), and Tumour Necrosis Factor(TNF) are well known in the art, and may be found in a number ofpublications, and patents. Representative suitable assays for use hereinare described in Adams et al., U.S. Pat. No. 5,593,992, whose disclosureis incorporated by reference in its entirety.

Interleukin -1 (IL-1)

Human peripheral blood monocytes are isolated and purified from eitherfresh blood preparations from volunteer donors, or from blood bank buffycoats, according to the procedure of Colotta et al, J Immunol, 132, 936(1984). These monocytes 1×10⁶) are plated in 24-well plates at aconcentration of 1-2 million/ml per well. The cells are allowed toadhere for 2 hours, after which time non-adherent cells are removed bygentle washing. Test compounds are then added to the cells for 1h beforethe addition of lipopolysaccharide (50 ng/ml), and the cultures areincubated at 37° C. for an additional 24h. At the end of this period,culture supernatants are removed and clarified of cells and all debris.Culture supernatants are then immediately assayed for IL-1 biologicalactivity, either by the method of Simon et al., J. Immunol. Methods, 84,85, (1985) (based on ability of IL-1 to stimulate a Interleukin 2producing cell line (EL-4) to secrete IL-2, in concert with A23187ionophore) or the method of Lee et al., J. ImmunoTherapy, 6 (1), 1-12(1990) (ELISA assay).

In Vivo TNF Assay:

(1) Griswold et al., Drugs Under Exp. and Clinical Res., XIX(6), 243-248(1993); or

(2) Boehm, et al., Journal Of Medicinal Chemistry 39, 3929-3937 (1996)whose disclosures

are incorporated by reference herein in their entirety.

LPS-induced TNFα Production in Mice and Rats

In order to evaluate in vivo inhibition of LPS-induced TNFα productionin rodents, both mice and rats are injected with LPS.

Mouse Method

Male Balb/c mice from Charles River Laboratories are pretreated (30minutes) with compound or vehicle. After the 30 min. pretreat time, themice are given LPS (lipopolysaccharide from Esherichia coli Serotype055-85, Sigma Chemical Co., St Louis, Mo.) 25 ug/mouse in 25 ulphosphate buffered saline (pH 7.0) intraperitoneally. Two hours laterthe mice are killed by CO₂ inhalation and blood samples are collected byexsanguination into heparinized blood collection tubes and stored onice. The blood samples are centrifuged and the plasma collected andstored at −20° C. until assayed for TNFα by ELISA.

Rat Method

Male Lewis rats from Charles River Laboratories. are pretreated atvarious times with compound or vehicle. After a determined pretreattime, the rats are given LPS (lipopolysaccharide from Esherichia coliSerotype 055-85, Sigma Chemical Co., St Louis, Mo.) 3.0 mg/kgintraperitoneally. The rats are killed by CO₂ inhalation and heparinizedwhole blood is collected from each rat by cardiac puncture 90 minutesafter the LPS injection. The blood samples are centrifuged and theplasma collected for analysis by ELISA for TNFα levels.

ELISA Method

TNFα levels were measured using a sandwich ELISA, as described inOlivera et al., Circ. Shock, 37, 301-306, (1992), whose disclosure isincorporated by reference in its entirety herein, using a hamstermonoclonal antimurine TNFα (Genzyme, Boston, Mass.) as the captureantibody and a polyclonal rabbit antimurine TNFa (Genzyme) as the secondantibody. For detection, a peroxidase-conjugated goat antirabbitantibody (Pierce, Rockford, Ill.) was added, followed by a substrate forperoxidase (1 mg/ml orthophenylenediamine with 1% urea peroxide). TNFαlevels in the plasma samples from each animal were calculated from astandard curve generated with recombinant murine TNFα (Genzyme).

LPS-Stimulated Cytokine Production in Human Whole Blood

Assay: Test compound concentrations were prepared at 10× concentrationsand LPS prepared at 1 ug/mI (final conc. of 50 ng/ml LPS) and added in50 uL volumes to 1.5 mL eppendorf tubes. Heparinized human whole bloodwas obtained from healthy volunteers and was dispensed into eppendorftubes containing compounds and LPS in 0.4 mL volumes and the tubesincubated at 37 C. Following a 4 hour incubation, the tubes werecentrifuged at 5000 rpm for 5 minutes in a TOMY microfuge, plasma waswithdrawn and frozen at −80 C.

Cytokine measurement: IL-I and/or TNF were quantified using astandardized ELISA technology. An in-house ELISA kit was used to detecthuman IL-1 and TNF. Concentrations of IL-1 or TNF were determined fromstandard curves of the appropriate cytokine and IC50 values for testcompound (concentration that inhibited 50% of LPS-stimulated cytokineproduction) were calculated by linear regression analysis.

CSBP/p38 Kinase Assay:

This assay measures the CSBP/p38-catalyzed transfer of ³²P from[a-³²P]ATP to threonine residue in an epidermal growth factor receptor(EGFR)-derived peptide (T669) with the following sequence:KRELVEPLTPSGEAPNQALLR (residues 661-681). (See Gallagher et al.,“Regulation of Stress Induced Cytokine Production by PyridinylImidazoles: Inhibition of CSBP Kinase”, BioOrganic & MedicinalChemistry, 1997, 5, 49-64).

Reactions were carried in round bottom 96 well plate (from Corning) in a30 ml volume. Reactions contained (in final concentration): 25 mM Hepes,pH 7.5; 8 mM MgCl₂; 0.17 mM ATP (the Km_([ATP]) of p38 (see Lee et al.,Nature 300, n72 pg. 639-746 (December 1994)); 2.5 uCi of [g-32P]ATP; 0.2mM sodium orthovanadate; 1 mM DTT; 0.1% BSA; 10% glycerol; 0.67 mM T669peptide; and 2-4 nM of yeast-expressed, activated and purified p38.Reactions were initiated by the addition of [gamma-32P]Mg/ATP, andincubated for 25 min. at 37° C. Inhibitors (dissolved in DMSO) wereincubated with the reaction mixture on ice for 30 minutes prior toadding the 32P-ATP. Final DMSO concentration was 0.16%. Reactions wereterminated by adding 10 ul of 0.3 M phosphoric acid, and phosphorylatedpeptide was isolated from the reactions by capturing it on p81phosphocellulose filters. Filters were washed with 75 mM phosphoricacids, and incorporated 32P was quantified using beta scintillationcounter. Under these conditions, the specific activity of p38 was400-450 pmol/pmol enzyme, and the activity was linear for up to 2 hoursof incubation. The kinase activity values were obtained aftersubtracting values generated in the absence of substrate which were10-15% of total values.

Representative final compounds of Formula (I) and (Ia) which have beentested, Examples 1 to 3, have all demonstrated positive inhibitoryactivity in this binding assay, having an IC₅₀ of <10 uM.

TNF-α in Traumatic Brain Injury Assay

This assay provides for examination of the expression of tumor necrosisfactor mRNA in specific brain regions which follow experimentallyinduced lateral fluid-percussion traumatic brain injury (TBI) in rats.Since TNF-α is able to induce nerve growth factor (NGF) and stimulatethe release of other cytokines from activated astrocytes, thispost-traumatic alteration in gene expression of TNF-α plays an importantrole in both the acute and regenerative response to CNS trauma. Asuitable assay may be found in WO 97/35856 whose disclosure isincorporated herein by reference.

CNS Injury Model for IL-b mRNA

This assay characterizes the regional expression of interleukin-1β(IL-1β) mRNA in specific brain regions following experimental lateralfluid-percussion traumatic brain injury (TBI) in rats. Results fromthese assays indicate that following TBI, the temporal expression ofIL-1β mRNA is regionally stimulated in specific brain regions. Theseregional changes in cytokines, such as IL-1β play a role in thepost-traumatic pathologic or regenerative sequelae of brain injury. Asuitable assay may be found in WO 97/35856 whose disclosure isincorporated herein by reference.

Angiogenesis Assay:

Described in WO 97/32583, whose disclosure is incorporated herein byreference, is an assay for determination of inflammatory angiogenesiswhich may be used to show that cytokine inhibition will stop the tissuedestruction of excessive or inappropriate proliferation of bloodvessels.

Rhinovirus/Influenza Assay:

Cell lines, rhinovirus serotype 39, and influenza virus A/PR/8/34 werepurchased from American Type Culture Collection (ATCC). BEAS-2B cellswere cultured according to instructions provided by ATCC using BEGM(bronchial epithelial growth media) purchased from Clonetics Corp. HELAcell cultures, used for detection and titration of virus, weremaintained in Eagle's minimum essential media containing 10% fetal calfserum, 2 mM 1-glutamine, and 10 mM HEPES buffer (MEM).

A modification of the method reported by Subauste et al., Supra, for invitro infection of human bronchial epithelial cells with rhinovirus wasused in these studies. BEAS-2B cells (2×10⁵/well) were cultured incollagen-coated wells for 24 hours prior to infection with rhinovirus.Rhinovirus serotype 39 was added to cell cultures for one hourincubation at 34° C. after which inoculum was replaced with fresh mediaand cultures were incubated for an additional 72 hours at 34° C.Supernatants collected at 72 hours post-infection were assayed forcytokine protein concentration by ELISA using commercially availablekits (R&D Systems). Virus yield was also determined from culturesupernatants using a microtitration assay in HELA cell cultures(Subauste et al., supra 1995). In cultures treated with p38 kinaseinhibitors, drug was added 30 minutes prior to infection. Stocks ofcompounds were prepared in DMSO (10 mM drug) and stored at −20° C.

For detection of p38 kinase, cultures were incubated in basal mediawithout growth factors and additives to reduce endogenous levels ofactivated p38 kinase. Cells were harvested at various timepoints afteraddition of rhinovirus. Detection of tyrosine phosphorylated p38 kinaseby immunoblot was analyzed by a commercially available kit and wasperformed according to the manufacturer's instructions (PhosphoPlus p38MAPK Antibody Kit: New England BioLabs Inc.).

In some experiments, BEAS-2B cells were infected with influenza virus(strain A/PR/8/34) in place of rhinovirus. Culture supernatant washarvested 48 and 72 hour post-infection and tested by ELISA for cytokineas described above.

Cells and Virus: Influenza A/PR/8/34 sub type H1N1 (VR-95 American TypeCulture Collection, Rockville, Md.) was grown in the allantoic cavity of10 day old chicken eggs. Following incubation at 37° C., andrefrigeration for 2½ hours at 4° C., allantoic fluid was harvested,pooled, and centrifuged (1,000 rcf; 15 min; 4° C.) to remove cells.Supernatent was aliquoted and stored at −70° C. The titer of the stockculture of virus was 1.0×10¹⁰ Tissue Culture Infective Dose/ml (TCID₅₀)

Inoculation procedure: Four-six week old female Balb/cAnNcrlBr mice wereobtained from Charles River, Raleigh, N.C. Animals were infectedintranasally. Mice were anesthetized by intraperitioneal injection ofKetamine (40 mg/kg; Fort Dodge Labs, Fort Dodge, Ia) and Xylazine (5mg/kg; Miles, Shawnee Mission, Ks) and then inoculated with 100 TCID50of PR8 diluted in PBS in 20 ul. Animals were observed daily for signs ofinfection. All animal studies were approved by SmithKline BeechamPharmaceuticals Institutional Animal Care and Use Committee.

Virus titration: At various times post infection, animals weresacrificed and lungs were aseptically harvested. Tissues werehomogenized, in vials containing 1 micron glass beads (Biospec Products,Bartlesville, Okla.) and 1 ml. of Eagles minimal essential medium. Celldebris was cleared by centrifugation at 1,000 rcf for 15 minutes at 4°C., and supernatants were serially diluted on Madin-Darby canine kidney(MDCK) cells. After 5 days of incubation at 37° C. (5% CO₂), 50 μl of0.5% chick red blood cells were added per well, and agglutination wasread after 1 hour at room temperature. The virus titer is expressed as50% tissue culture infective dose (TCID₅₀) calculated by logisticregression.

ELISA: Cytokine levels were measured by quantitative ELISA usingcommercially available kits. Ear samples were homogenized using a tissueminser in PBS. Cell debris was cleared by centrifugation at 14,000 rpmfor 5 minutes. The cytokine concentrations and thresholds weredetermined as described by the manufacturer; IL-6, IFN-γ, and KC (R&DSystems, Minneapolis, Minn.).

Myeloperoxidase Assay: Myeloperoxidase (MPO) activity was determinedkinetically as described by Bradley et al. (1982). Briefly, rabbitcornea were homogenized in Hexadecyl Trimethyl-Ammonium Bromide (HTAB)(Sigma Chemical Co. St. Louis, Mo.) which was dissolved in 0.5 mPotassium phosphate buffer (J. T. Baker Scientific, Phillipsburg, N.J.).Following homogenization, the samples were subjected tofreeze-thaw-sonication (Cole-Parmer 8853, Cole-Parmer, Vernon Hills, II)3 times. Suspensions were then cleared by centrifugation at 12,500×g for15 minutes at 4^(o)C. MPO enzymatic activity was determined bycolormetric change in absorbance during a reaction of O-Dianisidinedihydrochloride (ODI) 0.175 mg/ml (Sigma Chemical Co. St. Louis, Mo.)with 0.0002% Hydrogen peroxide (Sigma Chemical Co. St. Louis, Mo.).Measurements were performed by using a Beckman Du 640 Spectrophotometer(Fullerton, Calif.) fitted with a temperature control device. 50 ul ofmaterial to be assayed was added to 950 ul of ODI and change inabsorbance was measured at a wave length of 460 nm for 2 minutes at25^(o) C.

Whole Body Plethlysomography: Influenza virus infected mice were placedinto a whole body plethysomograph box with an internal volume ofapproximately 350-ml. A bias airflow of one 1/min was applied to the boxand flow changes were measured and recorded with a Buxco XA dataacquisition and respiratory analysis system (Buxco Electronics, Sharon,Conn.). Animals were allowed to acclimate to the plethysmograph box for2 min. before airflow data was recorded. Airway measurements werecalculated as Penh (enhanced pause). Penh has previously been shown asan index of airway obstruction and correlates with increasedintrapleural pressure. The algorithm for Penh calculation is as follows:Penh=[(expiratory time/relaxation time)-1] x (peak expiratory flow/peakinspiratory flow) where relaxation time is the amount of time requiredfor 70% of the tidal volume to be expired. Determination of arterialoxygen saturation. A Nonin veterinary hand held pulse oximeter 8500Vwith lingual sensor Nonin Medical, Inc., Plymouth Minn.) was used todetermine daily arterial oxygen saturation % SpO2 as described (Sidwellet al. 1992 Antimicrobial Agents and Chemotherapy 36:473-476).

Additional data and assay modifications may be found in PCT/US00/25386,(WO 01/19322) filed 15 Sep. 2000, whose disclosure is incorporatedherein by reference in its entirety.

Fluorescence Anisotropy Kinase Binding Assay

The CSBP kinase enzyme, a fluorescent ligand and a variableconcentration of the test compound are incubated together to reachthermodynamic equilibrium under conditions such that in the absence oftest compound the fluorescent ligand is significantly (>50%) enzymebound and in the presence of a sufficient concentration (>10×K_(i)) of apotent inhibitor the anisotropy of the unbound fluorescent ligand ismeasurably different from the bound value.

The concentration of kinase enzyme should preferably be ≧1×K_(f). Theconcentration of fluorescent ligand required will depend on theinstrumentation used, and the fluorescent and physicochemicalproperties. The concentration used must be lower than the concentrationof kinase enzyme, and preferably less than half the kinase enzymeconcentration.

A Typical Protocol is:

All components dissolved in Buffer of final composition 62.5 mM HEPES,pH 7.5, 1.25 mM CHAPS, 1.25 mM DTT, 12.5 mM MgCl₂ 3.3% DMSO.

p38 Enzyme concentration: 12 nM

Fluorescent ligand concentration: 5 nM

Test compound concentration: 0.1 nM-100 uM

Components incubated in 30 ul final volume in NUNC 384 well blackmicrotitre plate until equilibrium reached (5-30 mins)

Fluorescence anisotropy read in LJL Acquest.

Definitions: K_(i)=dissociation constant for inhibitor binding

-   -   K_(f)=dissociation constant for fluorescent ligand binding        The fluorescent ligand is the following compound:        which is derived from        5-[2-(4-aminomethylphenyl)-5-pyridin-4-yl-1H-imidazol-4-yl]-2-chlorophenol        and rhodamine green.

Representative final compounds of Formula (I) and (Ia) which have beentested, Examples 1 to 4, 6, 8 and 9, have all demonstrated positiveinhibitory activity in this binding assay, having an IC₅₀ of <1 uM.

SYNTHETIC EXAMPLES

The invention will now be described by reference to the followingexamples which are merely illustrative and are not to be construed as alimitation of the scope of the present invention. All temperatures aregiven in degrees centigrade, all solvents are highest available purityand all reactions run under anhydrous conditions in an Argon (Ar)atmosphere where necessary.

Mass spectra were run on an open access LC-MS system using electrosprayionization. LC conditions: 4.5% to 90% CH₃CN (0.02% TFA) in 3.2 min witha 0.4 min hold and 1.4 min re-equilibration; detection by MS, UV at 214nm, and a light scattering detector (ELS). Column: 1×40 mm Aquasil (C18)¹H-NMR (hereinafter “NMR”) spectra were recorded at 400 MHz using aBruker AM 400 spectrometer or a Bruker AVANCE 400. Multiplicitiesindicated are: s=singlet, d=doublet, t=triplet, q=quartet, m=multipletand br indicates a broad signal. For preparative (prep) hplc; ca 50 mgof the final products were injected in 500 uL of DMSO onto a 50×20 mm I.D. YMC CombiPrep ODS-A column at 20 mL/min with a 10 min gradient from10% CH₃CN (0.1% TFA) to 90% CH₃CN (0.1% TFA) in H₂O (0.1% TFA) and a 2min hold (unless otherwise stated). Flash chromatography was run overMerck Silica gel 60 (230-400 mesh) in solvent mixtures containingvarying relative concentrations of dichloromethane and methanol, orEtOAc, and hexane, unless otherwise stated. Chromatotron chromatographyas has been previously described (Desai, H K; Joshi, B S; Panu, A M;Pelletier, S W J. Chromatogr. 1985 223-227.) was run on chromatotronplates available from Analtech, Wilmington Del., USA.

satd=saturated; aq=aqueous; NMP=1-methyl-2-pyrrolidinone; soln=solution;other abbreviations are as described in the ACS Style Guide (AmericanChemical Society, Washington, D.C., 1986).

Example 1 8-(2,4,6-Trifluorophenyl)-4-(2,4-di-fluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

a)4-Chloro-6-(2,4,6-tri-fluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde

A solution of 4,6-dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde[Santilli, et al., J. Heterocycl. Chem. 1971, 8, 445-45] (1.0 gram(hereinafter “g”), 4.48 millimoles (hereinafter “mmol”)) in CHCl₃ (20milliliters (hereinafter “mL”)) was added 2,4,6-tri-fluoroaniline (0.735g, 5 mmol) followed by Et₃N (0.94 mL, 6.72 mmol, 1.5 equivalents(hereinafter “eq”)). The reaction mixture turned yellow and was heatedat 50⁰C for 8 hours (hereinafter “h”), 1 M aq Na₂CO₃ solution (50 mL)was added and the layers were separated. The organic layer was washedwith 50 ml sat'd. aqueous (hereinafter “satd aq”) NaCl solution, driedthrough anhydrous MgSO₄ and evaporated. The crude product was purifiedby flash chromatography (Hex/AcOEt=95/5) to afford 1.3 g (87%) of pure4-chloro-6-(2,4,6-trifluoro-phenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde.

LC MS (m/e)=334.0 (MH+).

b)4-(2,4,6-trifluorophenylamino)-6-(2,4-di-fluorophenyl)-2-methylsulfanyl-pyrimidine-5-carbaldehyde

To the product of the preceding example of step (a) (1.3 g, 3.9 mmol) indioxane (45 mL) and HO (15 mL) was added anhydrous K₂CO₃ (1.62 g, 11.7mmol) followed by phenylboronic acid (0.93 g, 5.86 mmol, 1.5 eq). Thereaction mixture was degassed by bubbling a stream of Ar through thesolution for 10 min and then tetrakis(triphenylphophine)-palladium(225.6 mg, 0.195 mmol, 0.05 eq) was added. The reaction mixture washeated under reflux for 20 h, cooled 23°, the layers were separated.EtOAc (100 mL), followed by satd aq NaCl solution (100 mL), was added,the organic layer was separated and dried (MgSO₄), filtered and theyellow solution was concentrated under reduced pressure. The residue waspurified by flash chromatography (Hex/AcOEt=95/5) to afford 1.1 g (67%yield) of the title compound. LC MS (m/e)=412.2 (MH+).

c)8-(2,4,6-Trifluoro-phenyl)-4-(2,4-difluorophenyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one

To a solution of the product of the preceding example, step (b) (1.1 g,2.67 mmol) in pyridine (10 mL) was added Ac₂O (10 mL) and the reactionmixture was heated under reflux for 64 hours, concentrated under reducedpressure and the residue was dissolved in EtOAc (200 mL), washed with 1M aq Na₂CO₃, and H₂O and satd aq NaCl, dried (MgSO₄), filtered andconcentrated under reduced pressure. The yellow residue was purified byflash chromatography (Hex/AcOEt=90/10) to afford pure4,8-bis-(2-fluoro-phenyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one(0.93 g, 80% yield). LC MS (m/e)=436.2 (MH+).

d)8-(2,4,6-Trifluoro-phenyl)-4-(2,4-difluorophenyl)-2-methylsulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one

The product of the preceding example, step (c) (0.93 g, 2.13 mmol) inCH₂Cl₂ (20 mL) was added 3-chloro-peroxybenzoic acid (0.96 g, 4.3 mmol,77% purity) and the reaction mixture was stirred 1 h at 23°, 0.75 mlMe₂S was added to quench the reaction. Then 1 M aq Na₂CO₃ (20 mL) wasadded, the layers were separated, and the organic layer was washed withH₂O, dried (MgSO₄) and concentrated under reduced pressure. The yellowresidue was purified by flash chromatography (Hex/AcOEt=60/40) to afford4,8-bis-(2-chloro-phenyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(0.95 g, 95% yield) to afford the title compound. LC MS (m/e)=468.0(MH+).

e)8-(2,4,6-Trifluoro-phenyl)-4-(2,4-difluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

The product of the preceding example (0.95 g, 2.03 mmol) in1-methyl-2-pyrrolidinone (15 mL) was added serinol (0.925 g, 10.15 mmol)and the reaction mixture was stirred at 23°. After 14 h, H₂O (60 mL) wasadded, followed EtOAc (60 mL). The layers were separated. The organiclayer was washed with satd aq NaCl, dried (MgSO₄), filtered concentratedunder reduced pressure. The yellow residue was then purified by flashchromatography to afford (0.93 g, 96% yield) of title compound. LC MS(m/e)=479 (MH+) 1.72 (Rt, min).

Example 2 8-(2.4difluorophenyl)-4-(2,4-di-fluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

a)4-Chloro-6-(2,4-difluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde

To a solution of 4,6-dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde[Santilli, et al., J. Heterocycl. Chem. 1971, 8, 445-45] (4.0 g, 18.0mmol) in CHCl₃ (50 mL) was added 2,4-di-fluoroaniline (2.02 mL, 19.8mmol) followed by Et₃N (3.76 mL, 27 mmol, 1.5 eq). The reaction mixtureturned yellow and was heated to reflux for 6 h, H₂O (50 mL) was addedand the layers were separated. The organic layer was evaporated to give6.5 g (>100%) of the crude title compound which is pure enough to beused in the next step. LC MS (m/e)=316 (MH+).

b)4-(2,4-difluorophenylamino)-6-(2,4-di-fluorophenyl)-2-methylsulfanyl-pyrimidine-5-carbaldehyde

To the product of the preceding example, step (a) (5.67 g, 18 mmol) indioxane (150 mL) and H₂O (50 mL) was added anhydrous K₂CO₃ (7.47 g, 54mmol) followed by 2,4-difluorophenylboronic acid (3.41 g, 21.6 mmol, 1.2eq). The reaction mixture was degassed by bubbling a stream of Arthrough the solution for 10 min. and thentetrakis(triphenylphophine)-palladium (1.03 g, 0.90 mmol, 0.05 eq) wasadded. The reaction mixture was heated under reflux for 12 h, cooled to23°, the solvents were removed, EtOAc (400 mL), followed by H₂O (200mL), was added, the organic layer was separated. The organic phase waswashed with satd aq NaCl, dried (MgSO₄), filtered and the yellowsolution was concentrated under reduced pressure to afford 7.75 g (>100%crude yield) of the title compound which can be used directly in thenext step. LC MS (m/e)=394 (MH+).

c)8-(2,4-difluorophenyl)-4-(2,4-difluorophenyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one

To a solution of the product of the preceding example, step (b) (2.33 g,5.9 mmol) in pyridine (15 mL) was added Ac₂O (15 mL) and the reactionmixture was heated under reflux for 48 h, concentrated under reducedpressure and the residue was dissolved in EtOAc (200 mL), washed with 1M aq Na₂CO₃ twice, and H₂O and satd aq NaCl, dried (MgSO₄), filtered andconcentrated under reduced pressure. The yellow residue was purified byflash chromatography to afford the title compound (1.2 g, 48% yield forthree steps). LC MS (m/e)=418 (MH+).

d)8-(2,4-difluorophenyl)-4-(2,4-difluorophenyl)-2-methylsulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one

To the product of the preceding example, step (c) (2.0 g, 4.8 mmol) inCHCl₃ (60 mL) was added 3-chloro-peroxybenzoic acid (2.48 g, 14.3 mmol)and the reaction mixture was stirred 5 h at 23°, then 1 M aq Na₂CO₃ (100mL) was added, the layers were separated, and the organic layer waswashed with H₂O, dried (MgSO₄) and concentrated under reduced pressureto afford (2.02 g, 94% yield) of the title compound. LC MS (m/e)=450(MH+).

e)8-(2,4-difluorophenyl)-4-(2,4-difluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

To the product of the preceding example, step (d) (1.90 g, 4.3 mmol) in1-methyl-2-pyrrolidinone (60 mL) was added serinol (1.97 g, 21.7 mmol)and the reaction mixture was heated to 50°. After 1 h, H₂O (100 mL) wasadded, followed by Et₂O (100 mL) and EtOAc (100 mL). The layers wereseparated. The organic layer was washed with satd aq NaCl, dried(MgSO₄), filtered concentrated under reduced pressure. The yellowresidue was purified by Flash chromatography to afford (1.22 g, 62%yield) of title compound. LC MS (m/e)=461 (MH+) 1.62 (Rt, min).

Example 38-(2,4,6-Trifluorophenyl)-4-(2-fluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

a)4-Chloro-6-(2,4,6-trifluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde

A solution of 4,6-dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde[Santilli, et al., J. Heterocycl. Chem. 1971, 8, 445-45] (1.0 g, 4.48mmol) in CHCl₃ (20 mL) was added 2,4,6-trifluoroaniline (0.735 mg, 5mmol) followed by Et₃N (0.94 mL, 6.72 mmol, 1.5 eq). The reactionmixture turned yellow and was heated to 50° C. for 8 h, 1 M aq Na₂CO₃(50 mL) was added and the layers were separated. The organic layer waswashed with 50 mL satd aq NaCl solution, dried through anhydrous MgSO4and evaporated. The crude product was purified by flash chromatographyto afford 1.3 g (87%) of pure4-chloro-6-(2,4,6-trifluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde.LC MS (m/e)=334.0 (MH+).

b)4-(2,4,6-Trifluoro-phenylamino)-6-(2,4-di-fluorophenyl)-2-methylsulfanyl-pyrimidine-5-carbaldehyde

To the product of the preceding example, step (a) (0.89 g, 2.67 mmol) indioxane (30 mL) and H₂O (10 mL) was added anhydrous K₂CO₃ (1.11 g, 8.03mmol) followed by 2-fluorophenylboronic acid (0.56 g, 4.0 mmol, 1.5 eq).The reaction mixture was degassed by bubbling a stream of Ar through thesolution for 10 min and then tetrakis(triphenylphophine)-palladium (154mg, 0.134 mmol, 0.05 eq) was added. The reaction mixture was heatedunder reflux for 20 h, cooled to 23°, the layers were separated, EtOAc(100 mL), followed by satd aq NaCl solution (100 mL), was added, theorganic layer was separated and dried (MgSO₄), filtered and the yellowsolution was concentrated under reduced pressure. The residue waspurified by flash chromatography to afford 530 mg (50% yield) of thetitle compound. LC MS (m/e)=394.0 (MH+).

c)8-(2,4,6-Trifluoro-phenyl)-4-(2-fluorophenyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one

To a solution of the product of the preceding example, step (b) (530 mg,1.35 mmol) in pyridine (6 mL) was added Ac₂O (6 mL) and the reactionmixture was heated under reflux for 64 h, concentrated under reducedpressure and the residue was dissolved in EtOAc (100 mL), washed with 1M aq Na₂CO₃, and H₂O and satd aq NaCl, dried (MgSO₄), filtered andconcentrated under reduced pressure. The yellow residue was purified byflash chromatography to afford pure4,8-bis-(2-fluoro-phenyl)-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one(520 mg, 92% yield). LC MS (m/e)=418.2 (MH+).

d)8-(2,4,6-Trifluoro-phenyl)-4-(2-fluorophenyl)-2-methylsulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one

The product of the preceding example, step (c) (520 mg, 1.24 mmol) inCH₂Cl₂ (15 mL) was added 3-chloro-peroxybenzoic acid (555 mg, 2.48 mmol,77% purity) and the reaction mixture was stirred 1 h at 23°, 0.5 mL Me₂Swas added to quench the reaction. Then 1 M aq Na₂CO₃ (50 mL) was added,the layers were separated, and the organic layer was washed with H₂O,dried (MgSO₄) and concentrated under reduced pressure to afford4,8-bis-(2-chloro-phenyl)-2-methanesulfonyl-8H-pyrido[2,3-d]pyrimidin-7-one(470 mg, 84% yield) to afford the title compound. LC MS (m/e)=449.8(MH+).

e)8-(2,4,6-trifluorophenyl)-4-(2-fluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

The product of the preceding example, step (d) (135 mg, 0.3 mmol) in1-methyl-2-pyrrolidinone (2 mL) was added serinol (137 mg, 1.5 mmol) andthe reaction mixture was stirred at 23°. After 14 h, H₂O (30 mL) wasadded, followed by EtOAc (30 mL). The layers were separated. The organiclayer was washed with satd aq NaCl, dried (MgSO₄), filtered concentratedunder reduced pressure. The yellow residue was then purified by Flashchromatography to afford (120 mg, 87% yield) of title compound. LC MS(m/e)=461 (MH+) 1.65 (Rt, min).

Example 48-(2,4,6-Trifluorophenyl)-4-(4-fluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

This compound was prepared using the methods of example 1 herein, andsubstituting 4-fluorophenyl boronic acid in the procedure of example1(b) to afford the title compound. LC-MS: 461 (MH+, m/z), 1.69 (Rt,min).

Example 58-(2,4-difluorophenyl)-4-(2-fluorophenyl)-2-((S)-2-hydroxy-1-methylethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

This compound was prepared using the methods of example 2, andsubstituting 2-fluorophenyl boronic acid in the procedure of example2(b) and (S)-(+)-2-amino-1-propanol in the procedure of example 2(e) toafford the title compound. LC-MS: 427 (MH+, m/z), 1.82 (Rt, min).

Example 68-(4-fluorophenyl)-4-(2,4-difluorophenyl)-2-((S)-2-hydroxy-1-methylethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

This compound was prepared using the methods of example 2, andsubstituting 4-fluoroaniline in the procedure of example 2(a) and(S)-(+)-2-amino-1-propanol in the procedure of example 2(e) to affordthe title compound. LC-MS: 427 (MH+, m/z), 1.87 (Rt, min).

Example 78-(2,4-difluorophenyl)-4-(2,4-difluorophenyl)-2-((S)-2-hydroxy-1-methylethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

This compound was prepared using the methods of example 2, andsubstituting 2,4-difluoroaniline in the procedure of example 2(a), andsubstituting 2,4-di-fluorophenyl boronic acid in the procedure ofexample 2(b), and (S)-(+)-2-amino-1-propanol in the procedure of example2(e) to afford the title compound.

LC-MS: 445 (MH+, m/z), 1.95 (Rt, min).

Example 88-(2,4-Difluorophenyl)-4-(2-chloro-4-fluorophenyl)-2-(2-hydroxy-1-hydroxymethylethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

a) 2-chloro-4-fluorophenylboronic acid

To 1.05 g Mg in THF (20 mL) was added catalytic I₂ and then a 1 mLportion of the 2-Chloro-4-fluoro-iodobenzene (10 g total, 39 mmol) wasadded. The mixture was stirred, slowly warmed, and an exothermicreaction began. The remainder of the 2-Chloro-4-fluoro-iodobenzene wasadded at a rate to maintain the exotherm and then the reaction washeated to THF reflux for 2.5 h, then cooled to 23°. The resultingmixture was added with rapid stirring to a −70° solution of trimethylborate (4.53 mL, 40 mmol) in THF (40 mL). The reaction was stirred at<0° for 20 min and at 23° for 16 h, then poured into 2N HCl (150 mL),stirred 2 h, and the THF was removed under vacuum. The residual aqueoussolution was extracted with Et2O (3×200 mL) and the combined extractswere dried (Na₂SO4) and concentrated to afford a red solid. Triturationwith hexane and drying afforded the title compound as a tan solid. (1.89g, 28%). ¹H-NMR CD₃OD δ 7.09 (m, 1H), 7.21 (m, 1H), 7.38 (m, 1H).

b) 8-(2,4fluorophenyl)-4-(2-chloro-4-fluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

The title compound was prepared using the methods of example 2 andsubstituting the product of the preceding example in the procedure ofexample 2(b), to afford the title compound. LC-MS: 477 (MH+, m/z), 1.75(Rt, min).

Example 98-(2,3-difluorophenyl)-4-(2,4-difluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one

This compound was prepared using the methods of example 2 andsubstituting 2,3-difluoroaniline in the procedure of example 2(a), toafford the title compound. LC-MS: 461 (MH+, m/z), 1.67 (Rt, min).

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

The above description fully discloses the invention including preferredembodiments thereof. Modifications and improvements of the embodimentsspecifically disclosed herein are within the scope of the followingclaims. Without further elaboration, it is believed that one skilled inthe are can, using the preceding description, utilize the presentinvention to its fullest extent. Therefore, the Examples herein are tobe construed as merely illustrative and not a limitation of the scope ofthe present invention in any way. The embodiments of the invention inwhich an exclusive property or privilege is claimed are defined asfollows.

1. The compound which is: 8-(2,4,6-trifluorophenyl)-4-(2,4-difluorophenyl)-2-(2-hydroxy-1-hydroxymethylethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one; 4,8-Bis-(2,4-difluoro-phenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido-[2,3-d]-pyrimidin-7-one; 8-(2,4,6-trifluorophenyl)-4-(2-fluorophenyl)-2-(2-hydroxy-1-hydroxymethylethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one; 8-(2,4,6-trifluorophenyl)-4-(4-fluorophenyl)-2-(2-hydroxy-1-hydroxymethylethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one; 4-(2-fluoro-phenyl)-8-(2,4-difluoro-phenyl)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8H-pyrido-[2,3-d]-pyrimidin-7-one; 4-(2,4-Difluoro-phenyl)-8-(4-fluoro-phenyl)-2-((S)-2-hydroxy-1-methyl-ethylamino)-8H-pyrido-[2,3-d]-pyrimidin-7-one; 4,8-Bis-(2,4-difluoro-phenyl)-2-(2-hydroxy-1-methyl-ethylamino)-8H-pyrido-[2,3-d]-pyrimidin-7-one; 8-(2,4-Difluorophenyl)-4-(2-chloro-4-fluorophenyl)-2-(2-hydroxy-1-hydroxymethylethylamino)-8H-pyrido[2,3-d]pyrimidin-7-one; or 8-(2,3-difluorophenyl)-4-(2,4-difluorophenyl)-2-(2-hydroxy-1-hydroxymethyl-ethylamino)-8H-pyrido[2,3-d]pyrimidin-7- one; or a pharmaceutically acceptable salt thereof.
 2. A pharmaceutical composition comprising an effective amount of a compound according to claim 1 and a pharmaceutically acceptable carrier or diluent.
 3. A method of treating a CSBP/RK/p38 kinase mediated disease in a mammal in need thereof, which method comprises administering to said mammal an effective amount of a compound according to claim
 1. 4. The method according to claim 3 wherein the CSBP/RK/p38 kinase mediated disease is psoriatic arthritis, Reiter's syndrome, gout, traumatic arthritis, rubella arthritis, acute synovitis, rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and other arthritic condition, sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, cerebral malaria, meningitis, ischemic and hemorrhagic stroke, neurotrauma/closed head injury, asthma, adult respiratory distress syndrome, chronic pulmonary inflammatory disease, chronic obstructive pulmonary disease, silicosis, pulmonary sarcososis, bone resorption disease, osteoporosis, restenosis, cardiac and brain and renal reperfusion injury, congestive heart failure, coronary arterial bypass grafting (CABG) surgery, thrombosis, atheroschlerosis, glomerularnephritis, chronic renal failure, diabetes, diabetic retinopathy, macular degeneration, graft vs. host reaction, allograft rejection, inflammatory bowel disease, Crohn's disease, ulcerative colitis, neurodegenrative disease, muscle degeneration, diabetic retinopathy, macular degeneration, tumor growth and metastasis, angiogenic disease, influenza induced pneumonia, eczema, contact dermatitis, psoriasis, sunburn, or conjunctivitis.
 5. A method of treating the common cold or respiratory viral infection caused by human rhinovirus (HRV), other enteroviruses, coronavirus, influenza virus, parainfluenza virus, respiratory syncytial virus, or adenovirus in a human in need thereof which method comprises administering to said human an effective amount of a compound according to claim
 1. 6. The method according to claim 5 wherein the respiratory viral infection exacerbates asthma, exacerbates chronic bronchitis, exacerbates chronic obstructive pulmonary disease, exacerbates otitis media, exacerbates sinusitis, or wherein the respiratory viral infection is associated with a secondary bacterial infection, otitis media, sinusitis, or pneumonia.
 7. The compound which is: 4-(2,4-difluoro-phenylamino)-6-(2,4,6-trifluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde; 4-(2,4-difluoro-phenyl)-6-(2,4-difluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde; 4-(2-fluorophenyl)-6-(2,4,6-trifluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde; 4-(4-fluorophenyl)-6-(2,4,6-trifluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde; 4-(2-fluorophenyl)-6-(2,4-difluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde; 4-(2,4-Difluoro-phenyl)-6-(4-fluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde; 4-(2,4-difluoro-phenyl)-6-(2,4-difluorophenylamino)-methylsulfanyl-pyrimidine-5-carbaldehyde; 4-(2-chloro-4-fluorophenyl)-6-(2,4-difluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde; or 4-(2,4-difluorophenyl)-6-(2,3-difluorophenylamino)-2-methylsulfanyl-pyrimidine-5-carbaldehyde. 